Figure 3.

Treatment with erastin induces cyst-lining epithelial cell death and promotes cyst growth in Pkd1^RC/RC mice kidneys. (A) Representative images of kidneys from Pkd1^RC/RC mice treated with erastin or vehicle from 1 to 3 months and percent cystic area relative to total kidney section area was significantly increased in kidneys from 3 months old Pkd1^RC/RC mice treated with erastin versus vehicle. Data reflect all sections quantified for each condition (n=8 in erastin-treated group and n=8 in vehicle-treated group). Scale bars, 2 mm. Treatment with erastin increased KW/BW ratios and BUN levels in Pkd1^RC/RC mice compared with those in vehicle-treated control mice. (B) Erastin treatment induced cyst-lining epithelial cell death in kidneys from Pkd1^RC/RC mice as detected by TUNEL assay. Scale bars, 50 μm. (C) The expression of cleaved caspase-3 in kidneys from Pkd1^RC/RC mice treated with erastin or vehicle analyzed by Western blot analysis. (D) Representative images of immunostaining with cleaved caspase-3 antibody in kidney sections from 3-month-old WT and Pkd1^RC/RC mice treated with erastin or vehicle. (E) The levels of nonheme irons in kidneys from Pkd1^RC/RC mice treated with erastin (n=8) or vehicle (n=8). (F) Representative images of immunostaining with 4HNE antibody in kidney sections from 3-month- old WT and Pkd1^RC/RC mice treated with erastin or vehicle. (G) Western blot analysis of GPX4 expression in kidneys from Pkd1^RC/RC mice treated with erastin or vehicle. (H) Representative images of Ki67 staining indicated that cell proliferation was increased in Pkd1^RC/RC kidneys treated with erastin versus that in Pkd1^RC/RC kidneys treated with vehicle. Scale bars, 50 μm. Statistical data are presented as the mean±SEM.