Fig. 1: TMX1 suppresses alternative TAG accumulation.
From: Identification of an alternative triglyceride biosynthesis pathway
a, Schematic of a haploid genetic screen in DGAT DKO cells using the stain BODIPY 493/503. Low and high represent the 5% of cells with the lowest and highest fluorescent signal, respectively. b, Fishtail plot depicting genetic regulators of lipid droplets in a screen of DGAT DKO HAP1 cells. Significant positive and negative regulators are coloured light blue and orange, respectively. The mutational index (MI) represents the ratio of inactivating gene-trap mutations per gene recovered from each (high and low) population (see Methods for a complete description). c, Immunoblot of TMX1 levels in HAP1 cell lines. WB, western blot; PDI, protein disulfide isomerase; LDHA, lactate dehydrogenase A. d, Quantitative increase in lipid droplets (visualized by BODIPY 493/503) in TMX1-knockout (ΔTMX1) HAP1 cells, as measured by flow cytometry. e, Lipid droplets, visualized by BODIPY 665/676 (green in the overlay), in HAP1 cell lines, including a double-DGAT and TMX1 triple knockout (3KO). Blue, Hoechst 33342. Scale bar, 10 μm. f, Lipid droplets (visualized by BODIPY 665/676) in ∆TMX1 293T cells. Blue, Hoechst 33342. Scale bar, 10 μm. g, TLC of neutral lipids in HAP1 (left) and 293T (right) cell lines. Cells were pulsed with 50 µM oleic acid (OA) for 24 h where indicated. 293T cells were additionally treated with 10 µM (each) DGAT inhibitor (DGATi) where indicated. h, Quantification of the increase in TAG induced by TMX1 deletion in HAP1 and 293T cell lines, normalized to WT cells. Data are mean ± s.e.m. of n = 3 independent experiments (two-way ANOVA, Bonferroni correction; each cell line was analysed separately).
