Scaffolding domain of CAV1 is crucial for inhibition of CLIC endocytosis.

<p>CAV1−/− MEFs were transiently transfected with (A) CAV1G83S-mCherry, (B) RFP-CAV3 and Cav3G55S-HA (C) CAV1Δ80-100-HA and (D) CAV1-SD-GFP (GFP-tagged minimal caveolin scaffolding domain) respectively. (A–D) Internalization assay was performed with anti-CD44 mAb and Tfn-647 for 2 min at 37°C. Cells were acid washed prior to fixation and internalized anti-CD44 mAb was labeled with AF-555 secondary antibody. 40–50 cells from each transfection were quantified for normalized fluorescence intensity of endocytic markers. In (A–D) data represent mean ± SEM of three independent experiments. **p = 0.002,***p = 0.0007, ****p<0.0001 (two-tailed t-test).