Figure 6. VPS13A decorates Lipid droplets.

(A) HEK293T cells were transfected with VPS13A-GFP for 24 hr and Lipidtox red was used as a marker for LDs. (B) HEK293T cells transfected with VPS13A-GFP for 48 hr were pulsed with 1 μM BODIPY-FA (red) at 37°C for 30 min followed by a chase in medium containing 500 uM OA for 2 hr at 37°C. (C) A close-up image of a LD in a cell taken from B in vivo is shown. Line profile analysis across the LD showed the enrichment of the VPS13A-GFP signal on the periphery of the LD (C’). Scale bar = 1 μm. Scale bars = 10 μm (A, B) and 1 μm (C).

Figure 6—figure supplement 1. Endogenous VPS13A is enriched at fractions containing LDs upon OA induction Workflow of LDs isolation and sucrose gradient fractionation.

FBS starved HeLa cells were processed or were subsequently incubated for 24 hr with 500 µM OA in FBS free medium, lysed and fractionated in 5–30% sucrose density gradients. Proteins in collected fractions were concentrated by TCA precipitation and subsequently separated by SDS- PAGE.