Fig. 3: Mogat1 inhibition suppresses lipid droplet formation.
From: Mogat1 drives metabolic adaptations to evade immune surveillance
a Schematic of the glycerol phosphate pathway and the monoacylglycerol pathway for triacylglycerol (TAG) synthesis. Key enzymes are indicated: GPAT (glycerol-3-phosphate acyltransferase), AGPAT (1-acylglycerol-3-phosphate acyltransferase), PAP (phosphatidic acid phosphatase), DGAT (diacylglycerol acyltransferase), MGAT (monoacylglycerol acyltransferase). b Confocal microscopy analysis of Mogat1 localization in MDA-MB-231 cells overexpressing Mogat1 tagged with neogreen (Mogat1-neogreen). Cells were stained for endoplasmic reticulum (ER) markers Calnexin and Calreticulin. Scale bar: 10 μm. c Western Blot of Mogat1 knockdown efficiency and Mogat1 overexpression in MMTV-PyMT and MDA-MB231 tumor cells. Quantification of d diacylglycerol (DAG) and e triacylglycerol (TG) content in MMTV-PyMT and MDA-MB231 tumor cells. (n = 4 biological cell cultures in data d, n = 3 biological cell cultures in data e). f Analysis of lipid droplet content in MMTV-PyMT and MDA-MB231 tumor cells. Representative images of Oil Red O staining and quantification of Oil Red O staining intensity. Scale bar: 20 μm. (n = 6 biological cell cultures). g Representative confocal microscopy images of lipid droplets in MMTV-PyMT and MDA-MB231 tumor cells following Mogat1 knockdown and mogat1 overexpression. Lipid droplets are visualized with BODIPY 493/503 (green). Scale bar: 20 μm. (n = 6 biological cell cultures). h–j Confocal microscopy analysis of lipid droplet marker Perilipin 2 (PLIN2) in h MDA-MB231 and i MMTV-PyMT cells. j Quantification of lipid droplet marker Perilipin 2. Scale bar: 20 μm. (n = 6 biological cell cultures). k, l Flow cytometry analysis of BODIPY 493/503 staining (a measure of neutral lipids) in MMTV-PyMT and MDA-MB231 tumor cells. k Representative histograms. l Mean fluorescence intensity (MFI) values. (n = 4 biological cell cultures). m, n Assessment of fatty acid uptake in MMTV-PyMT and MDA-MB231 tumor cells with or without Mogat1 knockdown and mogat1 overexpression using BODIPY FL C16. m Representative histograms. n Quantification of BODIPY FL C16 uptake. (n = 4 biological cell cultures). Data in (d, e, f, g, j, l, n) are presented as the mean ± SD; Statistical significance was calculated using one-way ANOVA (d, e, f, g, j, l, n). ns, not significant. Representative immunoblots and gels shown in (c) were repeated at least two times independently with similar results. Source data are provided as a Source Data file.
