Fig. 1: SARS-CoV-2 nsp6 protein binds to LD membrane.

a HeLa cells were transfected with mCherry-nsp6 or mCherry control vector for 24 h, and then were treated with 200 μM OA for another 6 h. LDs were stained with Bodipy493/503. Scale bar, 10 μm. The quantitative analysis was represented on the right panel (n = 10 cells). b Huh-7 cells were transfected with mCherry-nsp6 or mCherry control vector for 24 h. LDs were stained with Bodipy493/503. Scale bar, 10 μm. The quantitative analysis was plotted on the right panel (n = 10 cells). c HeLa cells were co-transfected with GFP-PLIN2 and mCherry-nsp6 or mCherry control vector for 24 h, and then were treated with 200 μM OA for another 12 h. Cells were scanned by confocal microscopy. Scale bar, 10 μm. The quantitative analysis was represented on the right panel (n = 14 cells). d Huh-7 cells were transfected with mCherry-Ctrl or mCherry-nsp6, followed by immunofluorescence staining for endogenous PLIN2. Scale bar, 10 μm. The quantitative analysis was represented on the right panel (n = 12 cells). e HeLa cells co-transfected with GFP-PLIN2 and mCherry-nsp6 or mCherry control vector were treated with 200 μM OA for 12 h. LDs were stained with LipidTOX Deep Red. Scale bar, 10 μm. The quantitative analysis was represented on the right panel (n = 14 cells). f Immunogold electron micrographs of Huh-7 cells expressing GFP-Vector or GFP-nsp6 treated with 200 µM OA for 12 hours. Gold particles are indicated by black dots. Scale bar, 200 nm. Quantitative analysis is shown in the right panel (n = 14 cells). g HeLa cells were transfected with mCherry-nsp6 or mCherry control vector for 36 h. The S7 fraction of mCherry-nsp6 overexpressed cells were subjected to subcellular fractionation by sucrose density gradient centrifugation shown in the right panel. All data are representative of three independent experiments with similar results (a–g) and are shown as the mean ± SD (a–f); two-tailed unpaired Student’s t test. PNS, post-nuclear supernatant. S7, soluble fraction. See also Supplementary Fig. 1. Source data are provided with this paper.