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. 2022 Sep 1;24(9):1364–1377. doi: 10.1038/s41556-022-00974-0

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 3 — a, Depletion of ERES components, Rab, tethering-complex components, and SNAREs results in endogenous GPAT4 co-localizing with ER marker Halo-KDEL without enrichment around LDs. Confocal imaging of live EGFP-GPAT4 endogenous knock-in cells upon RNAi of membrane fusion machinery components, transiently transfected with Halo-KDEL construct, 20 h after 1 mM oleic acid treatment. LDs were stained with MDH. Representative images from 3 independent experiments are shown. Scale bar, 5 and 1 μm (inlay). b, Effect of Sec12, Sar1, Sec23, Rab1, Rint1, Syx5, and Bet1 depletion on LD targeting of GPAT4 is rescued by expressing wildtype proteins. Confocal imaging of live EGFP-GPAT4 endogenous knock-in cells upon RNAi of Sec12, Sar1, Sec23, Rab1, Rint1, Syx5, and Bet1, followed by mCherry (mC) or Halo tagged constructs, 20 hr after 1 mM oleic acid treatment. Scale bar, 5 and 1 μm (inlay). c, Bar graph showing LD targeting ratios from the imaging experiment in b. Mean ± SD, n = (left to right) 30, 29, 27, 26, 26, 26, 42, 54, 29, 25, 31, 24, 32, 31 cells examined over 3 independent experiments. Two-tailed Student’s t-test, #p < 0.0001, compared to respective control transfection. Source numerical data are available in source data.

Source data