Fig. 1: Characterizing the phenotype of the GDC0032-acquired -resistant cell lines.

A A scheme illustrating the transformation of sensitive cells to resistant by exposing cells to increasing concentrations of GDC0032, starting at 1 nM and increasing up to 1 uM (see Material & Methods section). B, C Proliferation of IGROV1 and OAW42 sensitive (black) and resistant (red) cells, treated with DMSO or 100 nM GDC0032 for 4 days. The graphs (C) Scatter plot represents the growth, monitored by a live cell imager every 24 h (mean ± S.E.M. n = 3, *P < 0.05, ***P < 0.001). After 4 days of incubation, cells were fixed and stained using crystal violet (B). D GDC0032 IC:50 values of IGROV1^Sen (3.5e-9M) and IGROV1^Res (8e-7M) or OAW42^Sen (4.2e-7M) and OAW42^Res cells (1.9e-6M). (n = 6) e FACS analysis of the population in S phase in all cell lines, 48 h after treatment with DMSO or 100 nM GDC0032 (mean ± S.E.M. n > = 4, *P < 0.05, ***P < 0.001). F Whole-cell lysate western blot analysis of AKT, S6, and ERK from both cell lines, sensitive and resistant, in a time series of 100 nM GDC0032 treatment, compared to the beta-actin level. G GDC0032 IC:50 values of IGROV1^Res and OAW42^Res with the AKT inhibitor, MK2206 compared to GDC0032 alone showed in a scatter plot (mean ± S.E.M. n = 3, ****P < 0.0001)