PKC activation and phosphorylation of EBP50 promotes microvillar rearrangement. (A) JEG-3 cells were serum starved overnight, treated with 1 µM PMA for 30 min, then fixed and stained for endogenous EBP50 (red), ezrin (green), and actin (blue). Bar, 5 µm. (B) GFP-EBP50 wild-type and PKC phospho-mimetic (S162,339,340D) or -deficient (S162,339,340A) mutants were overexpressed in cells treated with PMA, stained for ezrin (red) and actin (blue), and their apical surface phenotype was scored. Bar, 10 µm. (C) Results from scoring JEG-3 cells for their presence of microvilli clusters; error bars indicate mean ± SD.