Fig. 2: Inactivation of SmarcAL1 gene in cells disrupts cellular lipid homeostasis and SmarcAL1 KO in mice drastically increases plasma triglyceride (TG) levels.

A, B Inactivation of SmarcAL1 gene in cells induces massive lipid droplet (LD) formation. Heterologous (A) and homologous (B) inactivation of SmarcAL1 genes in McA (A) and Huh7 (B) cells, respectively, induced LD formation captured under light microscopy (top). SmarcAL1 expression was confirmed with Western analysis (three replicates for each cell line) with anti-SmarcAL1 and -β-actin antibodies (bottom). C1 and C4 in A represent two McA clones with heterologous inactivation of SmarcAL1 gene. C Confirmation of SmarcAL1 heterozygous (HET) or homozygous (KO) gene deletion in mice. Western analysis of SmarcAL1 expression of liver extracts from the mice as indicated using anti-SmarcAL1 and -GAPDH antibodies. See Figs. S10–S12 for full images of the Western blots. D Metabolomic analysis of plasma from the WT and KO littermates (3 mice each). Ratios were calculated by comparing the averages of each metabolite of KO mice with those of WT mice and minus one. E, F Inactivation of SmarcAL1 gene increases plasma TG levels in mice. Plasma TG levels from individual littermates of the WT, HET, and KO mice with normal chow diet. Individual plasma samples were measured for TG using TG colorimetric assay kit (Cayman). Each dot represents a single mouse (E). Comparison of the plasma TG and total cholesterol (TC) profiles from the littermates of the WT, HET, and KO mice. Pooled plasma samples (from six mice each) from WT, HET and KO mice were fractionated by FPLC, and the fractions were analyzed for TG and TC separately. TG-rich lipoprotein (TRL), LDL and HDL in the fractions are indicated (F). P values calculated from t test and two-way ANOVA as indicated.