Figure 3.

ANGPTL4 ASO reduces ANGPTL4 protein levels in mouse liver and adipose tissue. A: Schematic overview of the set-up of the study. Male C57BL/6J mice fed chow were randomly assigned to either treatment with Neg-Ctrl ASO (1.25 mg/kg, n = 23), low-dose ANGPTL4 ASO (0.625 mg/kg, n = 24), or high-dose ANGPTL4 ASO (1.25 mg/kg, n = 21) via subcutaneous injections twice a week for a total duration of 2 weeks. In each group, half of the mice were euthanized in the ad libitum fed state and half after a 24 h fast. B: Angptl4, Angptl3, and Angptl8 mRNA levels in the liver as determined by quantitative PCR (qPCR). C: Lpl and Lipc mRNA levels in the liver as determined by qPCR. D: Hepatic ANGPTL4 protein levels as determined by Western blot. Right panel: Livers from wild-type, Angptl4^−/−, and Angptl4-Tg mice were used as a reference. E: Hepatic LPL protein levels as determined by Western blot. Adipose tissue was used as a reference. L.E., long exposure; S.E., short exposure. F: Adipose tissue ANGPTL4 protein levels as determined by Western blot. Adipose tissue from wild-type, Angptl4^−/− and Angptl4-Tg mice was used as a reference. G: Adipose tissue LPL activity using EnzChek lipase substrate. H: Adipose tissue LPL protein levels as determined by Western blot. HSP90 was used as a loading control. In the graphs, the horizontal bar represents the mean and the error bars represent SEM. Asterisk indicates significantly different from Neg-Ctrl ASO according to Tukey’s post hoc test. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. HSP90, heat shock protein 90.