Figure 3.

PKCλ/ι deficiency influences the cell size and adhesion molecules of podocytes. A: Deficient and control podocytes were stained with an antibody against vinculin, and grayscale images were obtained using a Leica DMBL microscope (×40 magnification). Binary inverted images were constructed using ImageJ macros to obtain footprints of vinculin distribution. Measurement of cell area and number and size of focal adhesions was performed using ImageJ for PKCλ/ι-deficient cells (B) and PKCζ-deficient cells (C) compared with WT cells. Twenty high-power fields (×40 magnification) of each genotype were analyzed, and the experiments were performed three times in different cell clones. Scale bar = 50 μm (applies to all panels). D: Western blot analysis reveals significantly reduced levels of integrin β1 in PKCλ/ι^−/− podocytes and a nonsignificant reduction in vinculin in PKCλ/ι^−/− and PKCζ^−/− podocytes. Lysates were normalized to GAPDH. E: Quantification of at least three independent experiments of different cell clones. ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001.