Figure 4.

Trehalose mitigates triglyceride accumulation in multiple in vitro models of steatosis. A. Triglyceride (TG) quantification in hepatocytes treated with or without 5 mM fructose in the presence or absence of 100 mM trehalose. Data are shown as mean ± SEM for, N = 3 independent experiments, n = 3 replicates per treatment group per experiment. ***, P < 0.001 (one-way ANOVA and Sidak’s post hoc analysis). B. Quantitative, real-time RT-PCR (qPCR) analysis of carbohydrate response element binding protein (ChREBP) and glycerol phosphate acyltransferase (GPAT) mRNA in hepatocytes treated with fructose in the presence or absence of 100mM trehalose. Target abundances are normalized to α-actin expression within each sample. Data are shown as mean ± SEM for, N = 3 independent experiments, n = 3 replicates per treatment group per experiment. ****, P < 0.0001, ***, P < 0.001, and *, P < 0.05 (one-way ANOVA and Sidak’s post hoc analysis). C. TG quantification in hepatocytes treated with or without BSA-conjugated fatty acids (500μM) in the presence or absence of 100mM trehalose (48hr). Data are shown as mean ± SEM for, N = 3 independent experiments, n = 2–3 replicates per treatment group per experiment. ***, P < 0.001, (one-way ANOVA and Sidak’s post hoc analysis). D. TG quantification in microsomal triglyceride transfer protein-deficient primary hepatocytes treated with or without 100mM trehalose (48hr). ****, P < 0.0001 by 2-tail T-Test. Data are shown as mean ± SEM for, N = 3 independent experiments, n = 3 replicates per treatment group per experiment. E., TG quantification in WT and ATG16L1^HM hepatocytes treated with BSA alone (control) or with BSA-conjugated fatty acids in the presence or absence of trehalose (100mM, 48hr). Data are shown as mean ± SEM for, N = 3 independent experiments, n = 3 replicates per treatment group per experiment. ****, P < 0.0001 by one-way ANOVA and Sidak’s post-hoc test.