(A) Podocytes treated with PAN (25, 50, 75, 100, or 125 μg/ml) for 24 h showed a dose-dependent decrease in viability. (B, C) The protein expression of nephrin, a marker of podocyte injury, was decreased following PAN treatment at concentrations ≥50 μg/ml, demonstrating that a PAN concentration ≥50 μg/ml was sufficient to induce podocyte injury. (D, E) Podocytes were examined by microscopy at 0 and 24 h. After 24 h, podocytes that were treated with 75 μg/ml PAN migrated faster than the controls, suggesting that the motility of PAN-treated podocytes was abnormal. (F) Phalloidin and DAPI staining of untreated control podocytes showed fine F-actin stress fibers, whereas podocytes exposed to 75 μg/ml PAN show a loss of stress fibers and the appearance of a disordered and thick cortical stress fiber distribution. (*p<0.05 vs. control; **p<0.01 vs. control; ns, no statistical significance; n = 3. Black bar = 200 μm; White bar = 40 μm. PAN: Puromycin aminonucleoside)