Lipid induced fission of caveolae is counteracted by Dyn2-GFP. (A) Schematic of fusogenic liposome methodology. (B) Time series showing the incorporation of Bodipy-labeled cholesterol into the PM of a Cav1-mCh cell. Scale bar, 10 μm. (C) Quantification of track duration time in Cav1-mCh cells pre-treated with Ctrl or Dyn2 siRNA as indicated. Cells were non-treated (white) or treated with fusogenic liposomes containing cholesterol (black) for 20 min prior to imaging. Analyses were performed using Imaris software, and track mean from at least 13 cells per condition are shown ± SEM. (D) Quantification of track duration time in Cav1-mCh cells, Dyn2-GFP-Cav1-mCh cells, or GFP-EHD2-Cav1-mCh cells non-treated (white) or treated with fusogenic liposomes containing cholesterol (black) for 20 min prior to imaging. Analysis was performed using Imaris software, and track mean from at least 20 cells per condition are shown ± SEM. Significance was assessed using t test, **P ≤ 0.01, ***P ≤ 0.001.