Western Blot Bands Research Articles

RETRACTION: Nutritional Flavonoids Modulate Estrogen Receptor α Signaling.

F. Virgili , F. Acconcia , R. Ambra , A. Rinna , P. Totta , and M. Marino , "Nutritional Flavonoids Modulate Estrogen Receptor α Signaling," IUBMB Life 56, no. 3 (2004): 145-151, https://doi.org/10.1080/15216540410001685083. The above article, published online on 3 January 2008 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Efstathios S. Gonos; the International Union of Biochemistry and Molecular Biology; and Wiley Periodicals LLC. The retraction has been agreed due to concerns raised by third parties on the data presented in the article. Specifically, duplication of several Western Blot bands has been detected within Figure 2a. The relative raw data could no longer be accessed due to the considerable time elapsed since the article's publication. The corresponding author, Maria Marino, maintains that the concerns do not impact the overall results and conclusions of the study, and disagrees with the decision of retraction. However, the article is retracted as the editors have lost trust in the integrity of the data presented and consider the conclusions invalid.

RETRACTION: Pyrrolo[1,2-b][1,2,5]benzothiadiazepines (PBTDs) Exert Their Anti-proliferative Activity by Interfering With Akt-mTOR Signaling and bax:bcl-2 Ratio Modulation in Cells From Chronic Myeloid Leukemic Patients.

C. Di Stefano, G. Marfe, M. M. Trawinska, P. Sinibaldi-Salimei, R. Silvestri, S. Amadori and E. Abruzzese, "Pyrrolo[1,2-b][1,2,5]benzothiadiazepines (PBTDs) Exert Their Anti-proliferative Activity by Interfering With Akt-mTOR Signaling and bax:bcl-2 Ratio Modulation in Cells From Chronic Myeloid Leukemic Patients," Cancer Science 101, no. 4 (2010): 991-1000, https://doi.org/10.1111/j.1349-7006.2010.01490.x. The above article, published online on 07 January 2010 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Masanori Hatakeyama; the Japanese Cancer Association; and John Wiley & Sons Australia, Ltd. The retraction has been agreed due to several instances of duplications of western blot bands observed in Figures3, 4, 5 and 8. Furthermore, splicing and deletion of bands was uncovered in Figures4 and 7 respectively. Finally, western blot bands presented in Figure4 was also found published earlier in another article. The authors provided partial raw data but the inconsistencies found could not be resolved on this basis. Due to the extent and nature of the duplications, the editors consider the results and conclusions of this study invalid.

RETRACTION: Phytosomal-curcumin Antagonizes Cell Growth and Migration, Induced by Thrombin Through AMP-Kinase in Breast Cancer.

M. Hashemzehi, R. Behnam-Rassouli, S. M. Hassanian, M. Moradi-Binabaj, R. Moradi-Marjaneh, F. Rahmani, H. Fiuji, M. Jamili, M. Mirahmadi, N. Boromand, M. Piran, M. Jafari, A. Sahebkar, A. Avan, and M. Khazaei, "Phytosomal-curcumin Antagonizes Cell Growth and Migration, Induced by Thrombin Through AMP-Kinase in Breast Cancer," Journal of Cellular Biochemistry 119, no. 7 (2018): 5996-6007, https://doi.org/10.1002/jcb.26796. The above article, published online on 30 March 2018 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Christian Behl; and Wiley Periodicals LLC. The retraction has been agreed due to concerns raised by third parties on the data presented in the article. Specifically, possible duplication of Western Blot bands was identified within Figure 5. The raw data provided by the authors upon request did not address the concerns, as clear evidence of image manipulation and fabrication was detected. Consequently, the article is being retracted, as the editors have lost confidence in the integrity of the presented data and deem the conclusions invalid.

RETRACTION: Long Noncoding RNA WDFY3-AS2 Suppresses Tumor Progression by Acting As a Competing Endogenous RNA of MicroRNA-18a in Ovarian Cancer.

W. Li, S. Ma, X. Bai, W. Pan, L. Ai and W. Tan,"Long Noncoding RNA WDFY3-AS2 Suppresses Tumor Progression by Acting as a Competing Endogenous RNA of MicroRNA-18a in Ovarian Cancer," Journal of Cellular Physiology 235, no. 2 (2020): 1141-1154, https://doi.org/10.1002/jcp.29028. The above article, published online on 25 July 2019 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Alexander Hutchison; and Wiley Periodicals LLC. The retraction has been agreed due to several instances of duplications of Western Blot bands found between Figures 3j, 5k and 7i of this article and figures from another article published elsewhere in the same year by different authors. The authors were invited to comment on the concerns raised but did not respond. The editors consider the results and conclusion reported in this article unreliable.

Does the sonic hedgehog signaling pathway play a role in anti-reflux mechanism of bladder in children?

A hedgehog family ligand, namely, sonic hedgehog (SHH), was reported to be important in the development of bladder and ureter smooth muscle. In this prospective study, we aimed to determine protein expression of SHH in resected ureterovesical junction (UVJ) segments of children with vesicoureteral reflux (VUR). The study group included 19 children; 12 (63%) girls, 7 (37%) boys, who had ureteroneocystostomy operation; 3 (15.7%) right sided, 7 (36.8%) left sided, 9 (47.3%) bilateral, due to primary VUR between years 2015 and 2018. Totally, 28 UVJ segments were examined for Western Blot analysis to determine related protein expression levels. The mean Western blot band area of SHH gene pathway related protein was 3880.69 (2059.55-13941.61) while the mean area of β-Actin, the house-keeping gene, was 20180.25 (9530.39-26709.75) (p = 0.001). Correlation analyses between grade of reflux and protein expression of SHH gene pathways revealed no significant relation (p = 0.300). When the UV samples were grouped as low- and high-grade reflux and compared in terms of SHH protein expression levels, no statistically significant difference was found between groups (p = 0.818). We concluded that SHH signaling molecule which is effective in development of bladder and ureter smooth musculature might also be effective in etiopathology of reflux.

RETRACTION "Protective effects of N-acetyl cysteine on lipid peroxide metabolism on isoproterenol-induced myocardial infarcted rats,".

M. F. Nagoor Meeran and P. S. Mainzen Prince, Journal of Biochemical and Molecular Toxicology 25, no. 3 (2011): 151-157, https://doi.org/10.1002/jbt.20371. The above article, published online on 23 November 2010 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Hari K. Bhat; and Wiley Periodicals, LLC. The retraction has been agreed due to duplication of several Western blot bands observed in Figure 2. The authors did not provide an explanation or their raw data. The editors consider the results and conclusion reported in this article unreliable.

EXPRESSION OF CONCERN: Glycogen synthase kinase 3β (GSK3β) mediates 6-hydroxydopamine-induced neuronal death.

G. Chen, K. A. Bower, C. Ma, S. Fang, C. J. Thiele, and J. Luo, "Glycogen Synthase Kinase 3β (GSK3β) Mediates 6-Hydroxydopamine-induced Neuronal Death," The FASEB Journal 18, no. 10 (2004): 1162-1164, https://doi.org/10.1096/fj.04-1551fje. This Expression of Concern for the above article published online on May 07, 2004 in Wiley Online Library (wileyonlinelibrary.com), has been published by agreement between the journal Editor-in-Chief, Loren E. Wold; the Federation of American Societies for Experimental Biology; and Wiley Periodicals LLC. The Expression of Concern has been agreed following concerns raised regarding duplication of the western blot bands presented in Figures1a and 3a. Further, duplications of bands were also observed within Figure5c. Due to the length of time that has elapsed since the publication of this article, theoriginal data are no longer available to the authors. The journal has decided to issue an Expression of Concern to inform the readers of these concerns while the authors are repeating the experiments.

Prognostic Impact of High-Molecular-Weight von Willebrand Factor Multimer Ratio in Classical Low-Flow Low-Gradient Aortic Stenosis.

High-molecular-weight (HMW) von Willebrand factor (VWF) multimer deficiency occurs in classical low-flow, low-gradient (LF/LG) aortic stenosis (AS) due to shear force-induced proteolysis. The prognostic value of HMW VWF multimer deficiency is unknown. Therefore, we sought to evaluate the impact of HMW VWF multimer deficiency on clinical outcome. In this prospective research study, a total of 83 patients with classical LF/LG AS were included. All patients underwent dobutamine stress echocardiography to distinguish true-severe (TS) from pseudo-severe (PS) classical LF/LG AS. HMW VWF multimer ratio was calculated using densitometric Western blot band quantification. The primary endpoint was all-cause mortality. Mean age was 79 ± 9 years, and TS classical LF/LG AS was diagnosed in 73% (n = 61) and PS classical LF/LG AS in 27% (n = 22) of all patients. Forty-six patients underwent aortic valve replacement (AVR) and 37 were treated conservatively. During a mean follow-up of 27 ± 17 months, 47 deaths occurred. Major bleeding complications after AVR (10/46; 22%) were more common in patients with HMW VWF multimer ratio <1 (8/17; 47%) in comparison to patients with a normal multimer pattern (2/29; 7%) at baseline (p = 0.003). In a multivariable Cox regression analysis, HMW VWF multimer deficiency was a predictor of all-cause mortality (HR: 3.02 [95% CI: 1.31-6.96], p = 0.009) in the entire cohort. This association was driven by higher mortality rates in the AVR group (multivariable-adjusted HR: 9.4; 95% CI 2.0-43.4, p = 0.004). This is the first study to demonstrate the predictive value of HMW VWF multimer ratio for risk stratification in patients with classical LF/LG AS. HMW VWF multimer deficiency was associated with an increased risk of all-cause mortality and major bleeding complications after AVR.

Retracted] MicroRNA‑671‑5p inhibits cell proliferation, migration and invasion in non‑small cell lung cancer by targeting MFAP3L.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that the western blotting data shown in Fig. 4B and the Transwell cell invasion data shown in Figs. 2D and 4E were strikingly similar to data appearing in different form in other articles written by different authors at different research institutes that had already been published elsewhere prior to the submission of this paper to Molecular Medicine Reports (one of which has been retracted). Moreover, there appeared to be inappropriately edited western blot bands featured in Figs. 4 and 5. In view of the fact that certain of the abovementioned data had already apparently been published previously, the Editor of Molecular Medicine Reports has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they agreed with the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 25: 30, 2022; DOI: 10.3892/mmr.2021.12546].

Comparison of the efficiency of the Sec and Tat secretory pathways in the secretion of recombinant neurturin protein using de novo designed signal peptides

Since cytoplasmic expression of heterologous proteins with disulfide bonds leads to the formation of inclusion bodies in E. coli, periplasmic production is preferable. The N-terminal signal peptide attached to the secreted protein determines the type of secretory pathway through which the target protein is secreted; Sec, Tat, or SRP. The aim of this study was to design and compare two novel signal peptides for the secretion of recombinant neurturin (as a model) via the Sec and Tat pathways. For this purpose, we aligned the natural signal peptides from E. coli and Bacillus subtilis to identify the conserved amino acids and those with the highest repetition. The SignalP4.1 and TatP1.0 software were used to determine the secretion efficiency of the new signal peptides. The efficiency of new signal peptides was then evaluated and compared experimentally with two naturally used signal peptides. Quantitative analysis of Western blot bands showed that approximately 80% of the expressed neurturin was secreted into the periplasmic space by new signal peptides. Circular dichroism spectroscopy also confirmed the correct secondary structure of the secreted neurturin. In conclusion, these novel signal peptides can be used to secrete any other recombinant proteins to the periplasmic space of E. coli efficiently.

Abstract P3164: Podocalyxin, A Renal Podocyte Specific Protein Expression In Urine Exosome Acts As A Marker For Podocyte Injury During Sars-cov-2 Infection

COVID-19 attributed to SARS-CoV2 infection has been associated with cardiovascular disease and diabetes. COVID-19 has been associated with endothelial cell dysfunction (ECD), which is manifested by entities such as microvascular thrombosis. ECD is quite common in type 2 diabetes mellitus (T2DM) where the renal podocyte dysfunction is often an early manifestation of microvascular complication. We explored whether presence of hyperglycemia predisposes to increased SARS-CoV2 infection and whether SARS-CoV2 infection puts an individual at a higher risk of developing cardio-metabolic complications such as kidney disease (DKD), with podocyte damage. We evaluated albuminuria and podocyte protein in urine derived exosomes in urine exosomes from SARS-CoV2 patients at 10 days, 6 months and 12 months post SARS-CoV2 infection. Methods: Blood and Urine samples from SARS-CoV2 patients post-acute phase of infection were procured. Peripheral blood mononuclear cells (PBMNCs) and urine exosomes were isolated and podocyte protein marker Podocalyxin (PODXL) was identified by western blot analysis. Results: Podocyte specific proteins were examined in urine exosomal fraction such as Podocalyxin and Nephrin (normalized to CD9 exosomal protein band intensities) levels increased significantly compared to urine samples from type-2 diabetes subjects with documented evidence of nephropathy. Podocalyxin levels were significantly high at 12month time point (p=0.001; n=9) and Nephrin levels were noted to be high at 10week (p=0.01; n=8), 6months (p=0.04; n=14 ) and 12Months (p=0.001; n=6) time points. Podocalyxin and Nephrin obtained from urine exosome samples collected from non diabetic subjects who never had COVID-19, showed little or barely detectable corresponding western blot bands. Interestingly, there was no significant differences noted on urine albumin: creatinine (UAR) ratios between the two groups, indicating that urine exosomal protein estimation may be an ideal biomarker to monitor podocyte damage in early phases of CKD. Conclusion: A persistent high levels of podocyte specific proteins was noted in urinary exosomes even at 12 months post acute Covid which may be secondary to long-standing podocyte inflammation leading to long term renal damage.

Retracted] MicroRNA‑505 is downregulated in human osteosarcoma and regulates cell proliferation, migration and invasion.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that the control GAPDH western blotting bands shown in Fig.4H on p.496 were strikingly similar to data that were submitted for publication in advance of this article in different form by different authors at different research institutes [LiuF, BaiC and GuoZ: The prognostic value of osteopontin in limited‑stage small cell lung cancer patients and its mechanism. Oncotarget8: 70084‑70096, 2017]. A further independent investigation conducted in the Editorial Office revealed that other western blotting data were likely to have been shared in common, comparing between the two articles. Owing to the fact that the contentious data in the above article had already been submitted for publication prior to the submission of this article to Oncology Reports, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, it was admitted that the authors Feng Chang, Jian-Na Liu and Jun-Xin Lin did not initially provide their agreement to be authors on this paper; otherwise, the rest of the authors accepted the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [Oncology Reports 39: 491‑500, 2018; DOI: 10.3892/or.2017.6142].

Panax notoginseng saponins promote hair follicle growth in mice via Wnt/β-Catenin signaling pathway.

Panax notoginseng saponins (PNS), the active ingredients of the traditional Chinese medicine Panax notoginseng, have strong neuroprotective and anti-platelet aggregation effects. To investigate whether PNS can promote hair follicle growth in C57BL/6J mice, the optimal concentration of PNS was initially determined, followed by clarification of the mechanism underlying their effects. Twenty-five male C57BL/6J mice had the hair on a 2 × 3cm2 area of the dorsal skin shaved and were equally divided into five groups: control group, 5% minoxidil (MXD) group, and three PNS treatment groups [2% (10mg/kg), 4% (20 mg/kg), and 8% (40 mg/kg) PNS]. They were then intragastrically administered the corresponding drugs for 28 days. The effects of PNS on C57BL/6J mice were analyzed by subjecting their dorsal depilated skin samples to different assessments, including hematoxylin and eosin staining, immunohistochemistry, immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting (WB). The group with 8% PNS exhibited the largest number of hair follicles from 14 days onwards. Compared with the control group, the number of hair follicles increased significantly in the mice treated with 8% PNS and 5% MXD, which significantly increased in a PNS-dose-dependent manner. Immunohistochemistry and immunofluorescence results revealed that treatment with 8% PNS activated the metabolism of hair follicle cells, with them showing higher rates of proliferation and apoptosis than those in the normal group. In qRT-PCR and WB analysis, the expression of β-catenin, Wnt10b, and LEF1 was upregulated in the PNS and MDX groups compared with that in the control group. Examination of the WB bands revealed that the greatest inhibitory effect of Wnt5a occurred in mice in the 8% PNS group. PNS may promote the growth of hair follicles in mice, with 8% PNS demonstrating the strongest effect. The mechanism behind this may be related to the Wnt/β-catenin signaling pathway.

Ashwagandha and Its Active Ingredient, Withanolide A, Increase Phosphorylation of TrkB in Cultured Hippocampal Neurons

Aims: To primary rat embryonic hippocampal neurons in culture, ashwagandha or one of its active ingredients, withanolide A were added in the presence or absence of nutrient supplementation and then assayed for activity of the BDNF receptor, TrkB.
 Study Design: Primary hippocampal neurons were cultured and grown in nutrient-rich or nutrient-poor medium. Ashwagandha or withanolide A were then be added to both types of media with or without an inhibitor of TrkB or either the PI-3K or MAPK pathway.
 Place and Duration of Study: Department of Biological Sciences, California State University, Los Angeles, CA, USA, between July 2021 and August 2022.
 Methodology: Rat embryos were removed by cesarean section from mother rats at 18 days’ gestation and the hippocampi of the former dissected, plated into culture dishes, and treated with the appropriate drug(s) (see Study Design above). After 4 days, neurons were harvested for Western blotting. Optical density of Western blot bands were quantified and statistically analyzed in a 2-way ANOVA, using a level of statistical significance at P < .05.
 Results: Under normal conditions (with N2 supplement), ashwagandha, but not withanolide A, increased phospho-TrkB immunoreactivity when compared to the effects of vehicle (controls, F(11, 24) = 22.48, P < .001), although withanolide A did not quite reach statistical significance (P = .069) when compared to that of the controlled condition. Likewise, under nutrient-deprived conditions, both ashwagandha and withanolide A also increased phosphorylation of TrkB when compared to that of vehicle-nutrient-deprived conditions (P < .0001). The same results were obtained in the presence of inhibitors of TrkB itself and the PI-3K (ashwagandha, P < .001; withanolide A, P < .001) and MAPK (ashwagandha, P = .027; withanolide A, P = .045) pathways.
 Conclusion: Ashwagandha or withanolide A activates TrkB, in nutrient-deprived hippocampal neurons, underscoring its role in neuronal survival signaling.

Retraction: Silencing of SOX12 by shRNA suppresses migration, invasion and proliferation of breast cancer cells.

This Retraction follows an Expression of Concern relating to this article previously published by Portland Press. This article is being retracted from Bioscience Reports at the request of the authors, following receipt of a notification from a reader alerting the Editorial Office to similarities that exist between the Figure 5C Cyclin/BT474 western blot bands from this article and the Figure 5C B-Carwnin/HO-8910 bands from a paper by Cheng et al. 2015 (doi: 10.18632/oncotarget.4541). During our investigation, the GAPDH bands from figure 3D were also highlighted by a reader as being very similar to those found in Figure 1E from Guo et al. 2017 (https://doi.org/10.1038/s41598-017-01832-y) and Figure 6E of Zheng et al. 2020 (https://doi.org/10.1186/s13046-019-1504-5). The authors were unable to provide the requested original data and responded to our queries by repeating the experiment, providing the Editorial Office with the new data. However, the Editorial Board did not feel that an adequate explanation to the duplicated blots was provided and remain concerned about the lack of original data. The authors therefore wish to retract the article, and the Editor-in-Chief and Editorial Board agree with the retraction.

Expression of Concern: Silencing of SOX12 by shRNA suppresses migration, invasion and proliferation of breast cancer cells.

The Editorial Office has been made aware of potential issues surrounding the scientific validity of this paper, hence has issued an expression of concern to notify readers whilst the Editorial Office investigates. Some of the western blot bands for Figure 5C, specifically the Cyclin/BT474 bands, appear to be highly similar to the Figure 5C B-Carwnin/HO-8910 bands in a paper by Cheng et al. 2015 (doi: 10.18632/oncotarget.4541), which has no related authors.

Retraction: α-bisabolol enhances radiotherapy-induced apoptosis in endometrial cancer cells by reducing the effect of XIAP on inhibiting caspase-3.

This Retraction follows an Expression of Concern relating to this article previously published by Portland Press. This article is being retracted from Bioscience Reports at the request of the authors following receipt of a notification from a reader alerting the Editorial Office to western blot bands that appear in both this paper and another paper by different authors (doi: 10.1186/s12935-019-0985-9). The authors wish to retract the article. The Editor-in-Chief and Editorial Board agree with the retraction.

Retraction: MiR-16-5p inhibits breast cancer by reducing AKT3 to restrain NF-kB pathway.

This article is being retracted from Bioscience Reports at the request of the authors due to technical reasons that they feel unable to address. This Retraction also takes place following receipt of a notification from a reader alerting the Editorial Office to similarities in figure panels across other published papers with no common authors. Figure panels of concern include 2C (common images across papers), 4F (common western blot bands across papers that have been vertically flipped) and 5C/F (common images across papers). The authors are unable to correct the article and wish to retract the article. The Editor-in-Chief and Editorial Board agree with the Retraction.

Aanalysis of Western blot band patterns in HIV-1 antibody positive cases

Aanalysis of Western blot band patterns in HIV-1 antibody positive cases

Editor's Note: Collagen Prolyl Hydroxylases Are Essential for Breast Cancer Metastasis.

The editors are publishing this note to alert readers to a concern about this article (1). In Fig. 1B, the Western blot bands representing HIF-1α, HIF-2α, and actin were inadvertently also used in Fig. 1A of a 2012 Oncogene publication by the authors (2). The blots were presented to demonstrate knockdown efficiency in subclones that were used for studies in both papers.