Fig. 3

Treatment with AUY922 induced heat shock responses, cell apoptosis, ROS production and TRPM1 attenuation in vitro. a Viability of 661 W (blue line) and ARPE19 (red line) cells, after treatment with varying concentrations of AUY922 for 3 d. Data are presented as the mean ± s.e.m. The P values were determined by two-tailed Student’s t-test, **** P < 0.0001. n = 3. b Representative images of clonogenic growth assays for ARPE19 (top) and 661 W (bottom) cells treated with indicated concentrations of AUY922 for 10 d. Five hundred cells/well were seeded in 6-well plates. n = 3. c Functional annotation of differentially expressed proteins in AUY922-treated 661 W cells. d Representative western blot results of 661 W, Mel1617 and A375 cells treated with varying concentrations of AUY922 for 48 h. β-Actin was used as a loading control. n = 3. e Representative western blot results of ARPE19 (left) and Mel1617 (right) cells stably expressing 3xF-TRPM1 after treatment with varying concentrations of AUY922 for 24 h. Cells expressing an empty vector were served as controls. n = 3. f Representative images of H&E stain and IHC analysis for TRPM1, AKT (T-AKT), phospho-AKT (P-AKT) and Ki67 expression in A375 xenograft tumor samples from the AUY922 treatment experiment in Fig. 1b. Scale bar: 25 μm. n = 3