Figure 3.

HGF is a novel inducer of NEPHRIN and NEPH1 phosphorylation.A, NEPHRIN-FLAG and NEPH1-FLAG were coexpressed with HGF in HEK293 cells, and phosphorylation was analyzed using their respective phosphoantibodies. SHP2 was also phosphorylated in the presence of HGF. Data are presented as mean ± SEM, and p-values were calculated using a two-tailed Student’s t test. B, HGF-expressing HEK293 cells were grown on the Transwell filter and stable NEPH1- and NEPHRIN-expressing HEK293 cells were cultured on the plastic at the bottom of the well. NEPHRIN and NEPH1 phosphorylation was measured by lysing the cells in the well, and Western blot analysis was performed with their respective phosphoantibodies. Data are presented as mean ± SEM, and p-values were calculated using a two-tailed Student’s t test. ∗∗∗∗p ≤ 0.001. All experiments were performed in triplicate and repeated three times with similar results, and representative images of the results are presented in the figure.