Fig. 3: Biomarkers to predict sensitivity to INX-315.

A Volcano plot illustrating the differential gene expression comparing cluster 3 (CDK2 addicted) versus cluster 2 (CDK4 addicted) and cluster 3 versus cluster 6 as mentioned in Fig. 1. The X axis represents log₂fold change and Y axis represents -log₁₀P. The P value was calculated based on two-tailed unpaired student t-test (B) Differential effect of CCNE1 and CDK2 depletion on cell cycle proteins as determined by western blotting in MB157 and HCC1806 cell lines. Western blotting was done at two independent times (C) Impact of ectopic overexpression of P16INK4A in HCC1806 cells on the cellular response to INX-315 (nM) following 48 h treatment as determined by western blotting. Western blotting was done at two independent times (D) Live cell imaging to monitor the impact of CDKN2A depletion in KURAMOCHI cells following the treatment with INX-315 (50 nM). Error bars were determined based on mean and SD from triplicates. The experiment was done at three independent times (n = 3). (*** indicates p value < 0.0001 as determined by 2-way ANOVA comparing siNT+INX-315 and siCDKN2A+INX-315). E Biochemical analysis to determine impact of CDKN2A depletion on the cell cycle proteins in the absence and presence of INX-315 (50 nM). F Live cell imaging to monitor the growth of KURAMOCHI-WT, KURAMOCHI-K4, MB157-WT and MB157-K4 cells in the absence and presence of INX-315. Error bars were determined from mean and SD from triplicates. Experiments were done at n = 3 independent times. *** indicates p value < 0.0001 as determined by 2 way-ANOVA comparing WT + INX-315 and CDK4-OE + INX-315. G Differential effect of INX-315 on the cell cycle proteins from MB157-WT and its isogeneic counterpart, MB157-K4, which harbors ectopic overexpression of CDK4 after 48-hour exposure. Western blotting was done at two independent times (B, C, E & G). Source data are provided as a source data file.