FIGURE 5:

CAV1-P158 has a shortened half compared with WT CAV1 in reconstituted Cav1^–/– MEFs. (A) Untransfected Cav1^+/+ MEFs (“WT MEF”) or Cav1^–/– MEFs transiently transfected with WT CAV1, CAV1-P158, or CAV1-P132L were incubated in the continuous presence of 100 μg/ml CHX and samples were collected at the indicated timepoints. Whole-cell lysates were analyzed by SDS–PAGE/Western blot. Densitometry was performed on blots probed with an anti-Cav1 antibody to determine the levels of endogenous or transfected CAV1 at each sampling time. (B) As in A, except Cav1^–/– MEFs were cotransfected with either WT CAV1+WT CAV1 or WT CAV1+CAV1-P158. Densitometry was performed on blots probed with an anti-Cav1 antibody for cells cotransfected with WT CAV1 + WT CAV1. For cells cotransfected with HA-P158 + WT Myc-CAV1, densitometry was performed on blots probed with a myc or HA antibody. We note that in cells cotransfected with HA-CAV1 and Myc-CAV1, HA-CAV1 appeared to be preferentially expressed for unknown reasons. Blots and densitometry results are representative of two independent experiments.