Figure 1.
MM cells are selectively sensitive to CDK7 inhibition. (A) Primary cells from patients newly diagnosed with MM (n = 4), MM cell lines (n = 30), PHA-activated PBMCs (n = 9), and nontransformed human cell lines (GMO5756, IMR90, HEEpiC, and HS-5 cell lines) were treated with different concentrations of YKL-5-124 for 48 hours and assessed for cell viability using CellTiter-Glo (CTG). IC50 analysis was performed with GraphPad software. Data are shown as the mean value ± SD; ∗∗∗P < .001. (B) H929 and AMO1 cells were engineered with a dTAG epitope (dTAG-CDK7WT). Cell viability was measured in H929 dTAG-CDK7WT and AMO1 dTAG-CDK7WT cells after treatment with dTAG˅-1 by CTG and represented as fold change increase compared with time of seeding (T0). (C) Control and YKL-5-124–treated MM cells were subjected to global quantitative TMT-based proteomic and phosphoproteomic analyses. KSEA for prediction of kinase activity was applied to identify activated (green bars) and inhibited kinases (red bar) in the YKL-5-124–treated group compared with control cells. (D) Whole-cell lysates from H929 cells treated with several concentrations of YKL-5-124 for 24 hours were subjected to western blot (WB) analysis and probed with indicated antibodies, with GAPDH or tubulin as a loading control (left). The ratio of phosphorylated/total forms of indicated CDKs was analyzed with Image J software and represented as fold change from untreated cells. Mean values ± SD in 3 MM cell lines is shown in the graph (right). (E) Whole-cell lysates from H929 cells treated with several concentrations of YKL-5-124 for different times (1, 4, 6, and 16 hours) were subjected to WB analysis and probed with selected antibodies (upper). The ratio of phosphorylated/total RNA polymerase II was analyzed with Image J software and represented as fold change from untreated cells. Mean values ± SD in 3 MM cell lines is shown in the graph (lower). IC50, 50% inhibitory; KSEA, kinase-substrate enrichment analysis; PBMCs, peripheral blood mononuclear cells; PHA, phytohemagglutinin; SD, standard deviation; TMT, tandem mass tag; WT, wild type.

MM cells are selectively sensitive to CDK7 inhibition. (A) Primary cells from patients newly diagnosed with MM (n = 4), MM cell lines (n = 30), PHA-activated PBMCs (n = 9), and nontransformed human cell lines (GMO5756, IMR90, HEEpiC, and HS-5 cell lines) were treated with different concentrations of YKL-5-124 for 48 hours and assessed for cell viability using CellTiter-Glo (CTG). IC50 analysis was performed with GraphPad software. Data are shown as the mean value ± SD; ∗∗∗P < .001. (B) H929 and AMO1 cells were engineered with a dTAG epitope (dTAG-CDK7WT). Cell viability was measured in H929 dTAG-CDK7WT and AMO1 dTAG-CDK7WT cells after treatment with dTAG˅-1 by CTG and represented as fold change increase compared with time of seeding (T0). (C) Control and YKL-5-124–treated MM cells were subjected to global quantitative TMT-based proteomic and phosphoproteomic analyses. KSEA for prediction of kinase activity was applied to identify activated (green bars) and inhibited kinases (red bar) in the YKL-5-124–treated group compared with control cells. (D) Whole-cell lysates from H929 cells treated with several concentrations of YKL-5-124 for 24 hours were subjected to western blot (WB) analysis and probed with indicated antibodies, with GAPDH or tubulin as a loading control (left). The ratio of phosphorylated/total forms of indicated CDKs was analyzed with Image J software and represented as fold change from untreated cells. Mean values ± SD in 3 MM cell lines is shown in the graph (right). (E) Whole-cell lysates from H929 cells treated with several concentrations of YKL-5-124 for different times (1, 4, 6, and 16 hours) were subjected to WB analysis and probed with selected antibodies (upper). The ratio of phosphorylated/total RNA polymerase II was analyzed with Image J software and represented as fold change from untreated cells. Mean values ± SD in 3 MM cell lines is shown in the graph (lower). IC50, 50% inhibitory; KSEA, kinase-substrate enrichment analysis; PBMCs, peripheral blood mononuclear cells; PHA, phytohemagglutinin; SD, standard deviation; TMT, tandem mass tag; WT, wild type.

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