Fig. 8. The CC domain and PR region are important for cell migration. (A) Podocytes were transfected with two different concentrations of CMS siRNA or control siRNA in triplicate. After 48 hours cells were subjected to serum-stimulated Boyden chamber migration assay. Cell lysates were analyzed by western blotting for CMS and actin. (B) Podocytes expressing mutated forms of CMS display reduced migration. Cells stably expressing CMS, CMS CC, CMS PR or vector control were subjected to a serum-stimulated migration assay for 4 hours. Data represent the mean ± s.d. of triplicates from three independent experiments (n=3; right panel; ^*P<0.01; ^**P=0.056). (C) Transformed NIH 3T3 cells display reduced migration. Src^*3T3 cells expressing the indicated peptides were subjected to a serum-stimulated migration assay for 2 hours. Data represent the mean ± s.d. of triplicates from three independent experiments (n=3; ^*P<0.0001; ^**P<0.002). P values were calculated using Student's t test. Protein expression was determined by western blot analysis.