Figure 1.

Phenotype of PKCλ/ι- and PKCζ-deficient podocytes in vitro. A: Phenotype of deficient or control podocytes after 10 days of differentiation at 37°C. Images of live cells were obtained using a light microscope (Axio Observer.Z1, ×20 magnification). PKCλ/ι^−/− podocytes grow spindle-like and do not reach confluence, in contrast to control and PKCζ^−/− cells. Scale bar = 100 μm (applies to all panels). B: Migration of deficient and control podocytes recorded using a live microscope (Axio Observer.Z1, ×20 magnification). The observed cells are shown in green, and the covered distance is illustrated as red dots. Scale bar = 50 μm (applies to all panels). C: Distance covered from startup to 6 hours, measured using ImageJ software (Manual Tracking), was significantly increased in PKCλ/ι^−/− podocytes. n = 10 cells per genotype. D: Phalloidin staining reveals increased number of cells with rearranged cytoskeleton in PKCλ/ι^−/− podocytes, podocytes treated with aPKC pseudosubstrate (PS) or with PKCλ/ι siRNA, in comparison with WT podocytes. PKCζ^−/− podocytes showed no difference from WT cells. Scale bar = 50 µm (applies to all panels). E: Quantification of cells with rearranged cytoskeleton. Each treated cell line was compared with an appropriate control. Ten high-power fields (×20 magnification) containing 10 to 40 cells were analyzed from each genotype using a Leica DMBL microscope. F: Transfection with PKCλ/ι siRNA significantly reduced the amount of PKCλ/ι in WT podocytes. ^∗∗P < 0.01.