Figure 3 from INX-315, a Selective CDK2 Inhibitor, Induces Cell Cycle Arrest and Senescence in Solid Tumors

TIS after INX-315 treatment of CCNE1-amplified cancers. A, Left, representative images after staining OVCAR3 and MKN1 cells for beta-galactosidase activity after treatment with INX-315 (scale bar = 100 μm). Experiments performed with three technical replicates. Right, quantification of integrated beta-galactosidase signal per cell. Dashed line indicates cutoff used to define beta-galactosidase positivity, and numbers represent percentage of beta-galactosidase–positive cells. B, Left, representative images after staining OVCAR3 and MKN1 cells with DAPI and phalloidin after treatment with INX-315 (scale bar = 100 μm). Right, quantification of relative nuclear size based on DAPI staining. Experiments performed with three technical replicates. C, ssGSEA scores derived from RNA sequencing of OVCAR3 cells treatment with control or INX-315 300 nmol/L for seven days. Scores are calculated for four independent senescence-associated gene sets; three technical replicates. D, GSEA plots derived from RNA-sequencing performed on tumor tissue from experiments in Fig. 2D and E. Normalized enrichment score (NES) and q value (false discovery rate) were calculated as described in Methods (all P values were calculated using unpaired t tests, error bars represent SD). E, ssGSEA scores for four senescence-associated gene signatures were calculated from RNA-sequencing on OV5398 PDX tumor tissue. Heat map shows z-scores for these. Bar plot above shows mean z-score for each sample across all signatures. F, GSEA plots derived from RNA-sequencing performed on tumor tissue from experiments in Fig. 2D and E. NES and q value (false discovery rate) were calculated as described in Methods (all P values were calculated using unpaired t tests, error bars represent SD).