Fig. 1. Caveolin-1-GFP behaves like endogenous caveolin-1. CHO cells stably transfected with caveolin-1-GFP were grown for 2 days until confluent. (A) Triton X-100-insoluble caveolae were separated from soluble membranes on sucrose gradients and immunoblotted with anti-caveolin-1. Caveolin-1-GFP (Cav-GFP) co-fractionated with endogenous caveolin-1 (Caveolin-1). The intensity of the bands for the two proteins indicates that caveolin-1-GFP constitutes a minor amount of the total caveolin-1 pool. (B) Cells were incubated for 2.5 hours in the presence of ³H-palmitate to label acylated proteins. Caveolin-1 was immunoprecipitated, separated by PAGE and processed for autoradiography. Both the endogenous protein (caveolin-1) and the caveolin-1-GFP (cav-GFP) were covalently labeled with ³H-palmitate. (C) Caveolin was extracted from cells using a combination of Triton X-100 and deoxycholate to solubilize the caveolar membranes and then analyzed by velocity gradient sedimentation. Both the endogenous caveolin (caveolin-1) and caveolin-1-GFP (cav-GFP) form oligomers that migrate to the bottom of the gradient.

Return to article