Fig. 8: Enhanced efficacy of INX-315 in combination with CDK4/6 inhibition. | Nature Communications

Fig. 8: Enhanced efficacy of INX-315 in combination with CDK4/6 inhibition.

From: Discrete vulnerability to pharmacological CDK2 inhibition is governed by heterogeneity of the cancer cell cycle

Fig. 8

A Scatter plot analysis in 1222, 3226 and HCC1806 cell lines following a combinatorial drug screen. X axis indicates the efficacy of individual drugs from the library in combination with DMSO. Y axis indicates the efficacy of individual drugs in combination with INX-315. B Venn Diagram illustrating the total number of drugs from cell line that cooperatively inhibited cell proliferation in combination with INX-315. C Live cell imaging to monitor the proliferation of the indicated cell lines following the treatment with palbociclib in combination with INX-315. Error bars represent mean and SD from triplicates. Experiments were done at n = 3 independent times. ***p < 0.0001 as determined by 2-wat ANOVA, comparing the INX-315 and Palbo/INX-315 treated groups. D DrugZ analysis from MIA PaCa-2 and HCC1806 cells indicating the positively and negatively selected guides following the selection with palbociclib. E Bi-variate flow cytometry analysis on MIA PaCa-2 and HCC1806 cells to indicate the combined effect of palbociclib and INX-315 on the BrdU incorporation within the S -phase population of cells. The experiment was done at n = 3 biological replicates for MiaPaCa-2 cells and n = 2 biological replicates for HCC1806 cells. Stacked bar plots represent the population of cells at each phase of the cell cycle. Error bars were determined from mean and SD from n = 4 biological replicates for MiaPaCa-2 and n = 3 biological replicates for HCC1806 cells. Fraction of cells within S-phase population incorporating BrdU was quantified. Statistical analysis was carried out based on two-tailed student t-test comparing the INX-315-treated and concurrent -treated groups. The p value for MiaPaCa-2 is 0.017. F The synergistic interaction between Palbo and INX-315 is based on BrdU incorporation. The synergistic interaction was determined using Synergy Finder, which calculates the Bliss synergy score. G Biochemical analysis on the indicated proteins from MCF7, MIA PaCa-2 and HCC1806 cell lines following the treatment with palbociclib in combination with INX-315 up to 48 h. MCF7 cells were treated with palbo (50 nM) and INX-315 (100 nM). MIA PaCa-2 cells were treated with Palbo (250 nM) and INX-315 (250 nM). HCC1806 cells were treated with Palbo (500 nM) and INX-315 (500 nM). H Immunoblotting in MIA PaCa-2-WT and MIA PaCa-2-RB-del cell lines following the treatment with palbociclib (250 nM) in combination with INX-315 (250 nM) up to 48 h to determine the differential impact on cell cycle proteins. Western blotting was done at two independent times (G & H). Source data are provided as a source data file.

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