FIGURE 4.
Role of c-Cbl in agonist-induced 5-HT2AR trafficking. Serum-deprived HEK293 cells, transiently transfected with control SCR or c-Cbl siRNA for 72 h (A), or transiently transfected with SCR or c-Cbl siRNA for 72 h and 5-HT2AR-v5 for 24 h (B), were immunoblotted with anti-c-Cbl or -β-actin antibodies (A) or 180 min after a synchronized pulse of 1 μm 5-HT (B), fixed, stained with anti-5-HT2AR-v5 antibody (visualized with Alexa Fluor 568-conjugated secondary antibody (red)) and anti-c-Cbl antibody (visualized with Alexa Fluor 488-conjugated secondary antibody (green)), and analyzed by confocal microscopy. White asterisk shows position of c-Cbl-depleted cells. Data shown are representative of three independent experiments. Bar, 5 μm.