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. 2020 May 4;9:e55038. doi: 10.7554/eLife.55038

Figure 6. GSLs are internalized to the endosomal system independent of Cav1, while Chol is predominantly trafficked to lipid droplets.

Figure 6.

(A) Cav1-mCh HeLa cells expressing Rab5-BFP were incubated with Bodipy-labeled LacCer or Chol for 15 min. Individual channels are shown for selected areas (dotted box). (B) Colocalization of lipids with Rab5-positive structures after indicated time-points. (C) Quantification of Cav1-mCh localization to Rab5-BFP positive structures before (control) and after lipid addition. Statistical analysis: ns - non significant. (D) Cav1 siRNA-treated Cav1-mCh HeLa cells expressing Rab5-BFP after incubation with Bodipy-labeled LacCer or Chol for 15 min. High-magnification images of selected areas (dotted box) for each channel are shown. (E) Quantification of EE positive for lipids in cells treated with siRNA control or against Cav1. Cells were incubated with Bodipy-lipids for 15 min. (F) Representative immunoblots of Cav1-mCh HeLa cells treated with control siRNA or siRNA against Cav1. Clathrin HC served as loading control. (G) Cav1-mCh HeLa cells were incubated with Bodipy-lipids for 15 min, fixed and LDs were stained using LipidTOX-DR. (H) Colocalization of lipids to LDs. (I) Colocalization of lipids with LDs in cells depleted of Cav1 after 15 min. (B, C, E, H, I) n = 10, mean ± SEM. All scale bars, 5 μm.

Figure 6—source data 1. Excel file containing source data pertaining to Figure 6B,C,E,H and I.