Extended Data Fig. 9: Characterization of Lpcat3-null 10T1/2 adipocytes.
From: Dietary control of peripheral adipose storage capacity through membrane lipid remodelling
(a) TIDE analysis (left) and Western blot analysis of LPCAT3 protein levels (right) validating Lpcat3-KO in 10T1/2 clonal cell lines transfected with sgRNAs targeting either exon 1 (L3_E1) or exon 3 (L3_E3). Calnexin served as a loading control (b) qPCR analysis of adipogenic markers in Lpcat3KO adipocytes expressing LPCAT3WT or LPCAT3H374A and differentiated for 6 days (n = 4, 4). (c) Shotgun lipidomic analysis of a wild-type 10T1/2 clonal cell line differentiated in the presence of 0.1% DMSO (veh ctrl) or the LPCAT3 inhibitor (R)-HTS-3 (10 µM) for 5 days. (d) Shotgun lipidomic analysis, (e) TG content (n = 3 wells/group), and (f) Oil red-O staining of Lpcat3KO adipocytes reconstituted with GFP Ctrl or the indicated GFP-tagged LPCAT3 fusion proteins (day 7 of differentiation). Data are presented as mean ± SEM. ***P < 0.001 by Welch’s t-tests with FDR correction using the Benjamini, Krieger, and Yekutieli procedure (b); or one-way ANOVA with Tukey’s multiple comparisons test (e).
