Gene Expression In Response Research Articles

Replication-IDentifier links epigenetic and metabolic pathways to the replication stress response

Abstract Perturbation of DNA replication, for instance by hydroxyurea-dependent dNTP exhaustion, often leads to stalling or collapse of replication forks. This triggers a replication stress response that stabilizes these forks, activates cell cycle checkpoints, and induces expression of DNA damage response genes. While several factors are known to act in this response, the full repertoire of proteins involved remains largely elusive. Here, we develop Replication-IDentifier (Repli-ID), which allows for genome-wide identification of regulators of DNA replication in Saccharomyces cerevisiae. During Repli-ID, the replicative polymerase epsilon (Pol ε) is tracked at a barcoded origin of replication by chromatin immunoprecipitation (ChIP) coupled to next-generation sequencing of the barcode in thousands of hydroxyurea-treated yeast mutants. Using this approach, 423 genes that promote Pol ε binding at replication forks were uncovered, including LGE1 and ROX1. Mechanistically, we show that Lge1 affects replication initiation and/or fork stability by promoting Bre1-dependent H2B mono-ubiquitylation. Rox1 affects replication fork progression by regulating S-phase entry and checkpoint activation, hinging on cellular ceramide levels via transcriptional repression of SUR2. Thus, Repli-ID provides a unique resource for the identification and further characterization of factors and pathways involved in the cellular response to DNA replication perturbation.

Oxidative Stress in Kidney of Zebrafish due to Individual and Combined Exposure to Amoxicillin, Arsenic, and Fluoride: Involving Nrf2-Keap1-ARE Pathway.

Toxic manifestations of different antibiotics and metal compounds have been studied comprehensively in the last decades; however, their co-toxicity on aquatic organisms is poorly investigated. This study aimed to evaluate the oxidative stress imposed on zebrafish kidney tissue when exposed to amoxicillin (AMX, 10 μg/L) alone or in combination with 50 μg/L of As2O3 (equivalent to 37.87 μg/L of As) and/or 15 mg/L of NaF (equivalent to 6.8 mg/L of F) for 15 days. We observed increased levels of cellular ROS, MDA, and GSH along with increased activity of CAT enzyme in all the treated groups. Disrupted histoarchitecture, including degeneration of tubular cells, vacuolation, and necrotic spots, was indicative of oxidative damage. mRNA expression of stress responsive genes like nrf2, gpx1, hsp70, keap1, nqo1, cat, and ho1 corroborated the data. Translocation of Nrf2 from cytoplasm to nucleus and its subsequent expression was higher for all the treated groups. Moreover, the mixture effects of AMX + As + F were more severe than the other combinations, while unique exposure with AMX had minimum effects. Highlighting the involvement of the Nrf2-Keap1-ARE pathway, these findings make us aware of the synergistic response of AMX, As, and F in the ecosystem, putting forward a great threat to humankind.

Leaf Gene Expression in Response to Boron Toxicity in Native Maize (Zea mays L.) Cultivars

Leaf Gene Expression in Response to Boron Toxicity in Native Maize (Zea mays L.) Cultivars

Regulation of Gut Starvation Responses Through Drosophila NP3253 Neurons.

The "gut-brain axis," a bidirectional communication system between the gastrointestinal tract and the central nervous system, plays a crucial role in regulating complex physiological functions in response to nutrients, pathogens, and microbiota. However, the cellular and molecular mechanisms governing this regulation remain poorly understood. Using Drosophila melanogaster as a model organism, we previously identified NP3253 neurons, located in both the brain and gut, as key contributors to gut homeostasis during oral bacterial infection. Here, we found a novel role of NP3253 neurons in regulating starvation resistance. We observed that a subset of NP3253 neurons in the gut were activated during starvation. To investigate downstream effect, we conducted RNA-Seq analysis on the gut of adult flies with genetically silenced NP3253 neurons, comparing gene expression under starved and fed conditions. This analysis identified 26 genes differentially expressed in response to both starvation and NP3253 neuronal activity. Among these, CG12643, encoding an uncharacterized short peptide, was found to be essential for starvation resistance in the gut. Our findings demonstrate that NP3253 neurons modulate the gut gene expression in response to starvation, thereby supporting physiological adaptation to environmental stressors.

Proteomic Analysis of Midgut of Silkworm Reared on Artificial Diet and Mulberry Leaves and Functional Study of Three UGT Genes

There remains a significant gap in production performance and disease resistance between silkworms reared on artificial diets and those reared on mulberry leaves. This study aims to identify key differential proteins through proteomic analysis of the midgut of silkworms fed artificial diets compared to those fed mulberry leaves. Utilizing molecular docking technology, three anti-nutritional factors that consistently bind to the UGT40B4, UGT340C2, and UGT40A1 proteins were selected, and the differential expression of these UGT genes in response to various anti-nutritional factors was examined. The findings indicate that variations in feed significantly influence the expression of digestive, metabolic, and immune-related proteins within the silkworm midgut. Notably, the expression levels of the UGT40B4, UGT340C2, and UGT40A1 genes vary across different silkworm organs and developmental stages, reflecting their respective physiological roles. Furthermore, the effects of soybean isoflavone, tannic acid, and arabinoxylan on silkworm growth and cocoon quality were found to differ when these substances were incorporated into semi-synthetic feed. This research is anticipated to provide valuable insights for future studies on the role of UGT genes in the silkworm midgut and the formulation of artificial diets for silkworms.

Overexpression of miR-155 Modulates Interferon Response and Inhibits Viral Replication of IHNV and IPNV in EPC Cells.

Infections caused by RNA viruses, including infectious haematopoietic necrosis virus (IHNV) and infectious pancreatic necrosis virus (IPNV), result in substantial economic losses in the aquaculture industry. MicroRNAs (miRNAs), particularly miR-155, play crucial roles in regulating host immune responses and viral infections. In this study, we investigated the overexpression effect of miR-155 in Epithelioma papulosum cyprini (EPC) cells infected with IHNV and IPNV and analysed the mechanisms underlying these antiviral activities. EPC cells were transfected with miR-155 at 40 pmol and then infected with IHNV or IPNV at an MOI of 0.01. The cytopathogenic effect (CPE) was observed for 5 days post-infection. The cells transfected with miR-155 did not show any signs of CPE or exhibit any viral growth over time after infection with both viruses. Additionally, real-time qPCR of type I interferon-related immune genes (ISG15 and Mx1) showed upregulation at 0, 24, and 48 h.p.i in cells transfected with miR-155. At 48 h post-infection, the cells transfected with miR-155 did not show any bands of viral protein by western blot. Furthermore, the overexpression of miR-155 in EPC cells significantly enhanced the expression of interferon response genes by targeting BCL2 and CYLD and suppressed the viral replication through directly targeting viral genes, including the L gene of IHNV and the VP2 gene of IPNV. These findings elucidate the dual mechanism of miR-155's antiviral effect through immune modulation and direct viral gene targeting, offering insights for developing novel antiviral strategies in aquaculture.

Adaptive Divergence and Functional Convergence: The Evolution of Pulmonary Gene Expression in Amphibians of the Qingzang Plateau.

The Qingzang Plateau, with its harsh environmental conditions-low oxygen, high ultraviolet radiation and significant temperature fluctuations-demands specialised adaptations for survival. While genetic adaptations have been extensively studied, gene expression's role in amphibian adaptation to high elevations remains understudied. This study analysed pulmonary gene expression in 119 amphibians across the plateau to explore how genetic and environmental factors shape expression evolution. Transcriptomic analyses revealed significant interspecies variation, driven by environmental factors like temperature, oxygen levels, UVB radiation and precipitation. Principal Component and Mantel analyses found no significant correlation between gene expression divergence and genetic distance. Instead, species-specific traits and environmental pressures were pivotal in shaping expression patterns. PERMANOVA analysis showed environmental factors had varying impacts on species. For instance, Bufo gargarizans exhibited a strong gene expression response to multiple environmental factors, while Scutiger boulengeri was less influenced, reflecting diverse adaptive strategies. Functional enrichment analysis highlighted convergence in key biological processes, such as energy metabolism, apoptosis and autophagy, despite species-specific gene expression differences. These processes are critical for surviving the plateau's extremes. The findings suggest that gene expression evolution in amphibians on the Qingzang Plateau is shaped by both genetic diversity and environmental pressures. Although gene expression profiles vary, they converge on essential functions, offering insights into adaptation mechanisms in extreme environments.

Blueberry-derived exosome like nanovesicles carry RNA cargo into HIEC-6 cells and down-regulate LPS-induced inflammatory gene expression: A proof-of-concept study.

Exosome-like nanovesicles (ELNs) of food origin have received great attention in the last decade, due to the hypothesis that they contain bioactive molecules. ELNs purified from edible species have been shown to be protective and are able to regulate intestinal homeostasis. Despite ELNs being potential rising stars in modern healthy diets and biomedical applications, further research is needed to address underlying knowledge gaps, especially related to the specific molecular mechanism through which they exert their action. Here, we investigate the cellular uptake of blueberry-derived ELNs (B-ELNs) using a human stabilized intestinal cell line (HIEC-6) and assess the ability of B-ELNs to modulate the expression of inflammatory genes in response to lipopolysaccharide (LPS). Our findings show that B-ELNs are internalized by HIEC-6cells and transport labeled RNA cargo into them. Pretreatment with B-ELNs reduces LPS-induced ROS generation and cell viability loss, while modulating the expression of 28 inflammatory genes compared to control. Pathway analysis demonstrates their ability to suppress inflammatory responses triggered by LPS. In conclusion, our data indicate that B-ELNs are up taken by HIEC-6cells and can modulate inflammatory responses after LPS stimulation, suggesting a therapeutic potential. This study demonstrates the role of B-ELNs in regulating crucial biological processes, like anti-inflammatory responses, which could support intestinal health.

Possible role of autophagy in microbial volatile pollutant-induced starch degradation and expression of hypoxia responsive genes.

Autophagy is thought to be critically involved in the regulation of nutrient metabolism and gene expression. Nevertheless, little is known about its role in regulating starch metabolism and hypoxia responsive genes in plants exposed to microbial volatile pollutants. In the present study, we found that exposure of Arabidopsis to Enterobacter aerogene (E. aerogene) volatile pollutants induced autophagy, as indicated by autophagosome formation. The exposure also caused upregulation of autophagy-associated genes, such as ATGs, NBR1, ATI1, and ATG8e-regulating transcription factors. Additionally, exposure to E. aerogenes volatile pollutants induced starch degradation in the roots of Arabidopsis seedlings. Finally, we found that ATG7-deficiency negatively affected the expression of hypoxia-responsive genes (i.e HRE1, HRA1, and ADH1) and starch degradation induced by E. aerogenes volatile pollutants. Overall, our study reveals that microbial volatile pollutants can induce starch degradation and autophagy, which participates in the regulation of some hypoxia-responsive genes and starch metabolism. These findings help to define the role of autophagy in plant nutrient metabolism and regulation of gene expression under microbial volatile pollutant exposure. The insights gained may contribute to agricultural management when living organisms face challenges from microbial volatile pollutants.

Advancing plant science through precision 3D bioprinting: new tools for research and biotech applications.

The integration of 3D bioprinting into plant science and biotechnology is revolutionizing research and applications. While many high-throughput techniques have advanced plant biology, replicating the complex 3D organization and cellular environments of plant tissues remains a significant challenge. Traditional 2D culture systems fall short of capturing the necessary spatial context for accurate studies of cell behavior, gene expression, and tissue development. Additionally, the lack of precise simulation of plant microenvironments limits control over cellular interactions and responses to external stimuli. Recent advancements in 3D bioprinting address these limitations by allowing precise control over cell positioning and biomaterial arrangement, thereby better replicating natural plant environments. This enables more accurate studies of gene expression, developmental processes, and stress responses. The technology also enhances our ability to test genetic modifications and biotechnological interventions, advancing crop improvement, sustainable agriculture, and precision breeding. This review examines the current state of 3D bioprinting in plant science, discusses its limitations, and explores its potential to transform research and applications in the field.

Genome-wide identification, phylogeny, evolutionary expansion, and expression analyses of ABC gene family in Castanea mollissima under temperature stress.

The ATP-binding cassette (ABC) gene family comprises some of the most critical transporter proteins in plants, playing vital roles in maintaining cellular homeostasis and adapting to environmental changes. While ABC transporters have been extensively characterized in various plant species, their profile in C. mollissima remains less understood. In this study, 164 ABC genes were identified and characterized within the C. mollissima genome, and subsequently classified into eight subfamilies. Collinear analysis suggested that dispersed duplication was the primary mechanism driving the expansion of the CmABC gene family. The study also examined morphological and physiological changes in C. mollissima under temperature stress, highlighting significant decreases in photosynthetic indicators and SOD enzyme activity, while other indicators varied. Transcriptome analysis revealed distinct expression patterns of various CmABC genes under temperature stress, identifying CmABCG29c and CmABCB11e as key candidates for responding to temperature stress. This was based on their expression patterns, correlation with physiological indicators, and WGCNA analysis. The expression levels of CmABC genes measured in RT-qPCR experiments were consistent with those observed in RNA-seq analysis. This research provides a theoretical foundation for understanding the physiological and gene expression responses of C. mollissima to temperature stress.

Integrated analysis of methylome and transcriptome responses to exercise training in children with overweight/obesity.

We examined the effects of a 20-wk exercise intervention on whole blood genome-wide DNA methylation signature and its association with the exercise-induced changes in gene expression profiles in boys and girls with overweight/obesity (OW/OB). Twenty-three children (10.05 ± 1.39 yr, 56% girls) with OW/OB were randomized to either a 20-wk exercise intervention [exercise group (EG); n = 10; 4 boys/6 girls] or to usual lifestyle [control group (CG); n = 13; 6 boys/7 girls]. Whole blood genome-wide methylome (CpG sites) analysis using Infinium Methylation EPIC array and transcriptome analysis using RNA-seq (STRT2 protocol) were performed. Exercise-induced modifications in DNA methylation at 485 and 386 CpGs sites in boys and girls, respectively. These CpG sites are mapped to loci enriched in distinct gene pathways related to metabolic diseases, fatty acid metabolism, and immune function. In boys, changes in the DNA methylation of 87 CpG sites (18% of the 485 CpGs sites altered by exercise) were associated with changes in the gene expression levels of 51 genes also regulated by exercise. Among girls, changes in DNA methylation at 46 CpG sites (12% of the initial 386 significant CpGs) were associated with changes in the expression levels of 30 exercise-affected genes. Genes affected by exercise that were associated with DNA methylation are related to obesity, metabolic syndrome, and inflammation. Multiomics analysis of whole blood samples from children with OW/OB suggests that gene expression response to exercise may be modulated by DNA methylation and involve gene pathways related to metabolism and immune functions.NEW & NOTEWORTHY This study pioneers the exploration into the effects of exercise on whole blood genome-wide DNA methylation patterns and its association with changes in transcriptome profiles in children with overweight/obesity. Exercise potentially impacts molecular pathways involved in metabolism and immune functions in children with overweight/obesity (sex-specific responses) through the modification of epigenetic and transcriptomic profiles. Our preliminary results provide initial steps to understand better the molecular mechanisms underlying cardiometabolic benefits of exercise in children with overweight/obesity.

HOS15 impacts DIL9 protein stability during drought stress in Arabidopsis.

HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE 15 (HOS15) acts as a substrate receptor of E3 ligase complex, which plays a negative role in drought stress tolerance. However, whether and how HOS15 participates in controlling important transcriptional regulators remains largely unknown. Here, we report that HOS15 physically interacts with and tightly regulates DROUGHT-INDUCED LIKE 19 (DIL9) protein stability. Moreover, application of exogenous abscisic acid (ABA) stabilizes the interaction between DIL9 and HOS15, leading to ABA-induced proteasomal degradation of DIL9 by HOS15. Genetic analysis revealed that DIL9 functions downstream to HOS15 and that the drought tolerance of hos15-2 plants was impaired in dil9/hos15 double mutants. Notably, DIL9 is directly associated with the promoter regions of ABF transcription factors and facilitates their expression, which is pivotal in enhancing ABA-dependent drought tolerance. Collectively, these findings demonstrate that HOS15 consistently degrades DIL9 under normal condition, while stress (drought/ABA) promotes the DIL9 activity for binding to the promoter regions of ABFs and positively regulates their expression in response to dehydration.

Altered mitochondrial unfolded protein response and FGF21 secretion in MASLD progression and the effect of exercise intervention

A high-calorie diet and lack of exercise are the most important risk factors contributing to metabolic dysfunction-associated steatotic liver disease (MASLD) initiation and progression. The precise molecular mechanisms of mitochondrial function alteration during MASLD development remain to be fully elucidated. In this study, a total of 60 male C57BL/6J mice were maintained on a normal or amylin liver NASH (AMLN) diet for 6 or 10 weeks. Some of the mice were then subjected to voluntary wheel running, while the other mice were fed a normal or AMLN diet until 14 and 18 weeks. The results showed that hepatic lipid deposition and the PERK-eIF2α-ATF4 pathway were significantly increased with prolonged duration of AMLN diet. However, expression of mitochondrial unfolded protein response (UPRmt) genes and mitokine FGF21 secretion were significantly enhanced in the 14-week AMLN diet mice, but were markedly reduced with the excessive lipid deposition induced by longer AMLN diet. Additionally, the exercise intervention acts as a regulator to optimize UPRmt signal transduction and to enhance mitochondrial homeostasis by improving mitochondrial function, reversing the UPRmt activation pattern, and increasing FGF21 secretion, which plays a pivotal role in delaying the occurrence and development of MASLD.

Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) is a plant pathogenic bacterium that possesses complicated motility regulation pathways including a typical chemotaxis system. A significant portion of our understanding about the genes functioning in Pst DC3000 motility is based on comparison to other bacteria. This leaves uncertainty about whether gene functions are conserved, especially since specific regulatory modules can have opposite functions in sets of Pseudomonas. In this study, we used a competitive selection to enrich for mutants with altered swimming motility and used random barcode transposon-site sequencing (RB-TnSeq) to identify genes with significant roles in swimming motility. Besides many of the known or predicted chemotaxis and motility genes, our method identified PSPTO_0406 (dipA), PSPTO_1042 (chrR) and PSPTO_4229 (hypothetical protein) as novel motility regulators. PSPTO_0406 is a homolog of dipA, a known cyclic di-GMP degrading enzyme in P. aeruginosa. PSPTO_1042 is part of an extracytoplasmic sensing system that controls gene expression in response to reactive oxygen species, suggesting that PSPTO_1042 may function as part of a mechanism that enables Pst DC3000 to alter motility when encountering oxidative stressors. PSPTO_4229 encodes a protein containing an HD-related output domain (HDOD), but with no previously identified functions. We found that deletion and overexpression of PSPTO_4229 both reduce swimming motility, suggesting that its function is sensitive to expression level. We used the overexpression phenotype to screen for nonsense and missense mutants of PSPTO_4229 that no longer reduce swimming motility and found a pair of conserved arginine residues that are necessary for motility suppression. Together these results provide a global perspective on regulatory and structural genes controlling flagellar motility in Pst DC3000.

Recombinant SpTransformer proteins bind to specific sites on sea urchin phagocytes and modulate SpTransformer gene expression and immune responsiveness

IntroductionThe California purple sea urchin, Strongylocentrotus purpuratus, relies exclusively on an innate immune system to survive in its pathogen rich marine environment. Central to this defense is the SpTransformer (SpTrf) gene family that is unique to the euechinoid group of echinoderms. These genes were initially identified based on their striking upregulation in response to immune challenge. The SpTrf gene family encodes structurally similar proteins with a wide range of sequence diversity within and among individual sea urchins. A recombinant (r)SpTrf protein interacts specifically with a variety of non-self targets. Other rSpTrf proteins cross-linked to inert beads show distinct functions for cell binding and augmenting phagocytosis . However, whether the rSpTrf proteins bind to sea urchin phagocytes, and the cellular consequences of binding are largely unexplored. MethodsrSpTrf protein binding to, and responses by phagocytes was investigated by cytology, flow cytometry, binding competitions using In-cell ELISA, and gene expression analyses. ResultsSoluble rSpTrf proteins bind specifically and exclusively to both live and fixed polygonal and small phagocytes. The different rSpTrf proteins appear to bind shared receptor(s) or other form of cell surface binding site. The phagocyte response to bound rSpTrf proteins culminates in modulated expression of the SpTrf gene family as well as other immune-related genes. ConclusionsThese findings underscore the multifaceted and dynamic functions of SpTrf proteins within the innate immune system of the purple sea urchin. Their varied functions enable a robust immune response while also providing a unique modulatory mechanism by which response levels are controlled and adjusted to the level of the foreign threat.

Exposomics: a review of methodologies, applications, and future directions in molecular medicine.

The exposome is the measure of all the exposures of an individual in a lifetime and how those exposures relate to health. Exposomics is the emerging field of research to measure and study the totality of the exposome. Exposomics can assist with molecular medicine by furthering our understanding of how the exposome influences cellular and molecular processes such as gene expression, epigenetic modifications, metabolic pathways, and immune responses. These molecular alterations can aid as biomarkers for the diagnosis, disease prediction, early detection, and treatment and offering new avenues for personalized medicine. Advances in high throughput omics and other technologies as well as increased computational analytics is enabling comprehensive measurement and sophisticated analysis of the exposome to elucidate their cumulative and combined impacts on health, which can enable individuals, communities, and policymakers to create programs, policies, and protections that promote healthier environments and people. This review provides an overview of the potential role of exposomics in molecular medicine, covering its history, methodologies, current research and applications, and future directions.

The Arabidopsis eATP receptor, DORN1 and CNGC19 channel act in tandem to regulate plant defense upon Spodoptera litura herbivory.

Plants deploy cellular Ca2+ elevation as a signal for environmental stress signaling. Extracellular ATP (eATP) is released into the extracellular matrix when cells are wounded. DOES NOT RESPOND TO NUCLEOTIDES 1 (DORN1), a key legume-type lectin receptor, senses and binds eATP and activates Ca2+ signaling. No evidence directly links calcium-mediated eATP signaling to resistance against insect herbivores. Here, we report upregulation of DORN1 transcripts upon wounding and Spodoptera litura feeding in Arabidopsis. Loss-of-function of DORN1 resulted in increased S. litura feeding compared to that on wildtype. Plant immunity is compromised in dorn1 mutants as they show reduced S. litura oral secretion mediated Ca2+ elevation, jasmonic acid accumulation, and expression of jasmonate responsive genes. The herbivory-induced calcium channel, CYCLIC NUCLEOTIDE GATED CHANNEL 19 (CNGC19), co-expresses with DORN1. We found that eATP-induced Ca2+ elevation requires functional CNGC19. Loss-of-function of DORN1 and CNGC19 highly increased the susceptibility to S. litura, mediated by reduced accumulation of jasmonates. We also demonstrate a plausible interaction of CNGC19 with DORN1. The data implicate the role of damage-released eATP and its receptor DORN1 in herbivory-induced defense signaling. DORN1 together with the Ca2+ channel CNGC19 generate the eATP-induced Ca2+ elevation, leading to the activation of immune signaling.

TFII-I/GTF2I regulates globin gene expression and stress response in erythroid cells.

Transcription factor TFII-I/GTF2I is ubiquitously expressed and has been shown to play a role in the differentiation of hematopoietic cells and in the response to various cellular stressors. We previously demonstrated that TFII-I acts as a repressor of adult β-globin gene transcription and positively regulates expression of stress response proteins, including ATF3. Here we analyzed the function of TFII-I in TF-1 cells during erythroid differentiation and in response to cellular stress, including unfolded protein response, hypoxia, and oxidative stress. Ablation of TFII-I leads to mild changes in cell cycle and proliferation of TF-1 cells. Importantly, TFII-I deficiency increased expression of the adult β-globin gene with concomitant reduction in the expression of the fetal γ-globin genes during erythropoietin mediated erythroid differentiation of TF-1 cells. Furthermore, TFII-I regulates genes involved in stress response, including CHOP, Elongin A, ATF3, ATF4, and Grp78, and participates in the apoptotic response to stressors. In summary, the data provide further support for a role of TFII-I in stress response and in the regulation of globin genes.

Diverse microtubule-destabilizing drugs induce equivalent molecular pathway responses in endothelial cells.

Microtubule (MT)-destabilizing drugs, despite their potential for toxicity, are used to treat a surprising range of diseases, including cancers, inflammatory disorders, and parasitic infections. To investigate how drugs with apparently similar actions on MTs achieve diverse clinical effects, we compared the molecular pathways that are activated by three drugs with different clinical profiles, vinblastine, combretastatin A4, and plinabulin. All three elicited similar gene expression responses via Rho GTPase activation. This finding suggests that their distinct clinical effects are not caused by different effects on MTs, but rather by differences in drug transport, pharmacokinetics or tubulin isotype affinity. Our findings provide insights into how plinabulin might protect the bone marrow and may help medicinal chemists design MT drugs for new applications.