Additional file 6 of AUY922 induces retinal toxicity through attenuating TRPM1

Additional file 6: Figure S6. a Representative western blot results of ARPE19 (left) and A375 (right) cells expressing either scrambled shRNA or shRNAs specific for TRPM1. Quantification analysis was performed for three independent experiments. Data are presented as the mean ± s.e.m. The P values were determined by two-tailed Student’s t-test, * P < 0.05; ** P < 0.01; n.s. = not significant. n = 3. b Representative images of clonogenic growth assays in TRPM1-knockdown cells described in Figure S6a. Five hundred ARPE19 (top) and 3000 A375 (bottom) cells were seeded in 6-well plates for 10 d. n = 3. c Representative western blot results of ARPE19 cells expressing either an empty vector or 3xF-TRPM1. Quantification analysis was performed for three independent experiments. Data are presented as the mean ± s.e.m. Significances were determined by two tailed Student’s t-test, n.s. = not significant. n = 3. d Representative images of clonogenic growth assays in ARPE19 cells stably expressing an empty vector or 3xF-TRPM1 after treatment with the indicated concentrations of AUY922 for 10 d. Five hundred cells/well were seeded in 6-well plates. n = 3. e Representative flow cytometry plots for apoptosis analyses are shown. Cells stably expressing an empty vector or 3xF-TRPM1 were treated with the indicated concentrations of AUY922 for 24 h before apoptosis analyses were performed. n = 3. f ROS levels were measured in 661 W cells treated with AUY922 for 48 h. Cells were labeled with DCFDA (20 μM) and analyzed by flow cytometry. ROS levels are expressed as fold change. Data are presented as the mean ± s.e.m. The P values were determined by two-tailed Student’s t-test, ** P < 0.01. n = 3.