Figure 5.

CAR3 suppression leads to enhanced CHRN endocytosis. C2C12 cells were transiently transfected with the indicated plasmids or specific siRNA using Lipofectamine 3000. 48 h later, the cells were either lysed and subjected to SDS-PAGE and immunoblotted with CAR2 or CAR3 antibody (A); or incubated with CHRN antibody (mAb210) at 4°C for 1 h, switched to 37°C for different times to induce CHRN endocytosis, subjected to acidic washes, then fixed and analyzed with flow cytometry (B); or were lysed and subjected to SDS-PAGE and immunoblotted with CHRN antibody (C). (D) The band densitometry was quantified using ImageJ software. (E) C2C12 cells were lysed and followed by immunoprecipitation with the indicated antibody, and then blotted with the specified antibodies. Data are mean ± SEM of 3 independent experiments, *p < 0.05, between the CAR3 group and the control group; ^#p < 0.05, between the siCar3 group and the siScram group.