Fig. 2

The effect of overexpressed ANLN E841K on the phenotype of podocytes. a Gene expression quantification of ANLN in three groups of cells were tested by RT‒qPCR. b-c Representative WB plots and WB quantification showed that podocyte gene overexpression cell lines were successfully constructed. d Representative immunofluorescence images of phalloidin. Red stands for Phalloidin, green for GFP, and blue for DAPI. The cytoskeleton of the mutant group was clustered and arranged disorderly. e, g Representative pictures of three group cells at the same position on plate at 0, 12, 24, and 36 h, respectively. The cell migration speed of E841K group was significantly faster than that of the WT group. f, h Representative immunofluorescence images of endocytosis. Red stands for Dextran, green for GFP, and blue for DAPI. The fluorescence intensity of podocytes with dextran in E841K was significantly higher than the other two groups. i, j Representative pictures of Vector, WT and E841K cells adhering to the 6-well plate. The adhesion ability of E841K cells was significantly reduced compared with that of the WT cells. k-m ANLN E841K interacted with CD2AP. k Representative WB plots of protein samples from the three groups of cells. l Representative fluorescence images of the three group cells. Red stands for CD2AP, green for ANLN, and blue for DAPI. m WB quantification of Flag protein pulled down by CD2AP. The amount of Flag protein pulled down by CD2AP in the mutant group was significantly less than that in the WT group. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001