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. Author manuscript; available in PMC: 2023 Jul 1.
Published in final edited form as: Nature. 2022 Dec 7;613(7942):160–168. doi: 10.1038/s41586-022-05507-1

Extended Data Fig. 11 ∣. CLSTN3β associates with CIDE proteins and blocks LD fusion.

Extended Data Fig. 11 ∣

(a—d) Western blot analysis of CLSTN3β-Flag co-immunoprecipitation assays in transfected HEK293FT cells. a, CIDEC-HA, b, CIDEA-HA, CIDECα-HA, and CIDECβ-HA, c, RAB18-HA and CGI58-HA, d, HA-tagged CIDEC C-terminal region (CIDEC-C-HA). e, Co-localization of CLSTN3β-GFP and CIDEA-HA in transfected 3T3-L1 preadipocytes treated with OA. Scale bar = 5 μm. (f—h) Fluorescence recovery after photobleaching (FRAP)-based lipid exchange rate assay in transfected 3T3-L1 preadipocytes incubated with 200 μM OA and 1 μg/mL BODIPY 558/568 C12 fatty acid for 16 h. f, Calculated lipid exchange rates (n = 15, 14, 17 cells). Cells used for representative traces are highlighted in red. g, Representative mean optical intensity traces for one pair of LDs from each group. h, Representative images of FRAP-based lipid exchange rate assay. Yellow arrows point to bleached LDs. Bar plots show mean ± SEM. Each point represents a biological replicate. Two-sided ***P < 0.001 by ordinary one-way ANOVA with Tukey’s multiple comparisons test (f).