(A) Treatment of podocytes with 300 μg/ml PS resulted in disrupted actin (red) and loss of stress fibers. Under these conditions, podocytes were largely devoid of synaptopodin (green). ML204 treatment blocked synaptopodin loss and cytoskeletal disruption. (B) Quantification of cytoskeletal remodeling (defined as no visible actin fibers) showed that ML204 rescued podocytes in a dose-dependent manner (n = 90 images/condition). (C) Loss of synaptopodin abundance and cytoskeletal remodeling by 300 μg/ml PS was prevented by TRPC5 shRNA–mediated Trpc5 gene silencing. Scr, scrambled shRNA control. (D) Quantification of cytoskeletal remodeling showed that TRPC5 shRNA–mediated Trpc5 depletion rescued podocytes, in contrast to scrambled control. n = 90 images/condition (10 images × 3 repeats × 3 independent trials). (E) Cell lysates from podocytes treated with PS or PS+ML204 showed that synaptopodin abundance, attenuated by 300 μg/ml PS compared with PBS-treated controls, was rescued by 10 or 30 μM ML204 in a dose-dependent manner. (F) Quantification of 4 Western blots from 4 independent experiments showed that 30 μM ML204 rescued synaptopodin abundance for PS-mediated degradation. Treatment with 10 μM ML204 trended in the same direction, but did not achieve statistical significance. (G) Rac1 activity (Rac1-GTP) increased in podocytes treated with 300 μg/ml PS compared with PBS-treated controls; 30 μM ML204 abrogated this activation. (H) Quantification of 3 Western blots from 3 independent experiments showed that 30 μM ML204 significantly blocked PS-mediated Rac1 activation. Original magnification, ×400 (A and C). *P < 0.05, ****P < 0.00001, ANOVA.