FIGURE 2 from Inhibition of NEK2 Promotes Chemosensitivity and Reduces KSHV-positive Primary Effusion Lymphoma Burden

Abstract

<p>JH295 treatment results in death of PEL cells but has no impact on normal B cells. <b>A,</b> IC<sub>50</sub> curves of PEL cell lines treated with a range of JH295 concentrations and normalized to the DMSO control values. Curves were fitted using nonlinear regression and are connected by means. Data are four independent biological replicates, each performed in triplicate. See also <a href="#SMT1" target="_blank">Supplementary Table S1</a>. <b>B,</b> Quantification of PEL cell death via Trypan blue staining following JH295 treatment. Individual data points from three independent biological replicates are plotted as mean ± SD and analyzed using one-way ANOVA with Dunnett multiple comparisons. ****, <i>P</i> < 0.0001; **, <i>P</i> = 0.0003; *, <i>P</i> = 0.04. <b>C,</b> PEL cell viability via Trypan blue count following mock or JH295 treatment. Individual data points from three independent biological replicates are plotted as mean ± SD and analyzed using one-way ANOVA with Dunnett multiple comparisons. ****, <i>P</i> < 0.0001; **, <i>P</i> < 0.007. <b>D,</b> IC<sub>50</sub> curve of bulk PBMCs treated with JH295 and normalized to the DMSO control values, overlayed on the PEL data from A. Curves were fitted using nonlinear regression and are connected by means. Data are three individual biological replicates, each performed in triplicate, using two unique PBMC donors. <b>E,</b> Cell viability of PBMCs versus PEL cells treated with JH295, each normalized to the corresponding DMSO control values. Individual data points from two independent biological replicates using two unique PBMC donors and performed in triplicate are plotted as mean + SEM and analyzed using one-way ANOVA with Dunnett multiple comparisons. ****, <i>P</i> < 0.0001. <b>F,</b> Cell viability of purified primary B cells treated with either DMSO or JH295. Data representing fold change in luminescence relative to the DMSO control, are plotted as individual data points of three to six independent biological replicates using three to six unique cell donors, and analyzed using unpaired two-tailed <i>t</i> test. ns, not significant. See also <a href="#SMF1" target="_blank">Supplementary Fig. S1</a>. <b>G,</b> Cell viability of purified primary B cells following mock or LPS stimulation. Data are fold change in luminescence relative to mock-stimulated cells, are plotted as individual data points of three to six independent biological replicates using three-six unique cell donors, and analyzed using unpaired two-tailed <i>t</i> test. **, <i>P</i> = 0.0031. <b>H,</b> Cell viability of stimulated purified primary B cells treated with and without JH295. Data are fold change in luminescence relative to the LPS control, are plotted as individual data points of three to six independent biological replicates using three to six unique cell donors, and analyzed using unpaired two-tailed <i>t</i> test. ns, not significant.</p>