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. 2024 Jan 20;15:631. doi: 10.1038/s41467-024-44978-w

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a Co-immunoprecipitation analysis of M-Flag with endogenous STX18. b GST pull-down analysis of GST-M with STX18-His. c, d HeLa cells expressing the M-HA and STX18-Flag were treated with 200 μM OA for 6 h, then fixed and immunostained with anti-HA and anti-Flag. Cells were imaged by confocal microscopy. Number of LDs per cell was counted from 50 cells of three independent experiments. e, f HeLa cells expressing the RFP-GFP-Plin2 were transfected with M-HA and STX18-Flag for 24 h and were treated with 200 μM OA for 12 h, then fixed and immunostained with anti-HA and anti-Flag. Cells were imaged by confocal microscopy. Number of RFP + GFP- dots per cell was counted from 50 cells of three independent experiments. g The effect of M-HA on the interaction of STX18-Flag with ATG14, Beclin1 and Vps34 was detected by immunoprecipitation. h The effect of SARS-CoV-2 infection on the interaction of STX18-Flag with ATG14 was detected by immunoprecipitation. i Atg14 knockout Vero-E6 cells were transfected with indicated plasmids for 24 h and infected with SARS-CoV-2 for 24 h. Meanwhile cells were treated with 200 μM OA for 12 h. Cells were fixed and immunostained with anti-LC3. Cells were imaged by confocal microscopy. Number of colocalization of LC3/LDs per cell was counted from 25 cells of three independent experiments. j, k Vero-E6 cells (j) or Atg14 knockout Vero-E6 cells (k) were transfected with indicated plasmids for 24 h and infected with SARS-CoV-2 for 24 h. Meanwhile cells were treated with 200 μM OA for 12 h. Cell lysates were analyzed via western blot. Viral RNA level was determined by RT-qPCR. Three independent experiments. Data information: LDs were labeled with LipidTOX Red (c) or BODIPY-493/503 (i). Scar bar represents 10 μm for all fluorescence images. Error bars, mean ± SD of three independent experiments. Two-tailed Unpaired Student’s t test. Source data are provided as a Source Data file.