Figure 5

CORO2B in human podocytes augments susceptibility to cellular stress. (a–c) Expression of CORO2B in human podocytes leads to a decrease in mean focal adhesion number per cell. Focal adhesions were visualized by PAXILLIN immunofluorescence staining. (n = 12 control and 12 CORO2B cells of one representative experiment were analyzed; >3 independent experiments were performed; ***p > 0.001), (d–e) Increased CORO2B expression enhances migratory capability of podocytes in single cell migration assays (n = 117 control and 100 CORO2B cells out of 3 independent experiments; **p < 0.01). (f–j) CORO2B expression results in a decrease of central actin cytoskeleton structures. F-ACTIN was visualized by Phalloidin staining. (n = 30 control and 36 CORO2B cells of one representative experiment were analyzed for central stress fibers (SF), >3 independent experiments were performed; ****p > 0.0001; cell type fraction was analyzed in 3 independent experiments by phenotyping of 100 cells per condition and experiment, data are significant (p < 0.001) for difference in intact central FA and AC group.) (k–m) CORO2B increases susceptibility to Doxorubicin induced cellular spreading defects. Cell spreading was performed on collagen IV coated glass cover slips for 1 hour. Podocytes were continuously treated with 1 µg/ml Doxorubicin for 24 hours before and during cell spreading assay (n = 329 V. ctrl., 327 CORO ctrl., 356 V. Doxo., 312 CORO. Doxo., cells of 3 independent experiments were analyzed per condition; ****P > 0.0001).