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. 2018 Dec 3;129(1):281–295. doi: 10.1172/JCI122595

Figure 3. Increased levels of phospholipids and chylomicron-associated proteins in Lpin2/3-KO intestine.

Figure 3

(A) Immunoblot analysis of intestinal protein levels for apolipoproteins apoB48, apoA-I, and apoA-IV, lipid droplet protein perilipin 2, phospholipid synthetic enzyme CCTα, and ER protein calnexin. Mice were fed a high-fat diet for 6 days. WT, wild-type. All panels are the same protein samples. The top 4 blots were run contemporaneously, with α-tubulin as a normalization control. The lower 4 blots were run contemporaneously with a nonspecific protein band as a normalizing control. Three biological replicates of each genotype are shown, and are representative of 5–8 samples of each genotype. (B–D) Proximal intestinal lipidomics analysis by electrospray ionization–tandem mass spectrometry in mice maintained on chow diet or fed a high-fat diet (HFD) for 6 days. Average ± SD, n = 4–6. (E) Altered PC composition in Lpin2/3-KO intestine, with reduced proportion of arachidonyl-PC species compared with WT. Average ± SD, n = 4–6. (F) CCTα (Pcyt) mRNA levels in intestine from mice indicated. Average ± SD, n = 4–6. (G) Immunoblot analysis of the mTORC1 target, p70S6 kinase, in intestine. Total and phosphorylated p70S6 kinase (Thr 389) were detected by specific antibodies. Blots were run contemporaneously with the same protein samples. *P < 0.05; **P < 0.01 by ANOVA.