Figure 2. Mboat7 knockdown promotes hepatic steatosis and worsen liver injury.

(A and B) C57BL/6 mice were fed chow or HFD with concurrent Control and Mboat7 ASO Injections at 12.5 mg/kg week for 20 weeks. Liver Mboat7 expression was measured via qPCR (A); n = 8–10; *p≤0.05, ****p≤0.0001; Two-way ANOVA with Tukey’s post-hoc test). The inset show a representative Western blot for hepatic MBOAT7 protein levels, which was replicated in n = 4 mice. (B) Body weight was measured weekly. (C) Food intake was measured in C57BL/6 mice at 12 weeks of diet and ASO injections (n = 6). (D) Oxygen consumption was measured by indirect calorimetry in C57BL/6 mice after 12 weeks of diet and ASO injections (n = 6). (E) Representative liver hematoxylin and eosin stained sections. 20x magnification. (F) Liver-to-body weight measurements from mice fed Chow and HFD with Control and Mboat7 ASO Injections for 20 weeks (n = 8–10; ****p≤0.0001; Two-way ANOVA with Tukey’s post-hoc test). (G and H) Plasma ALT (G) and AST (H) levels were measured after 20 weeks of diet feeding and ASO injections (n = 8–10; ****p≤0.0001; Two-way ANOVA with Tukey’s post-hoc test). (I) Percent steatosis quantified by a blinded board certified pathologist at the Cleveland Clinic (n = 3; ****p≤0.0001; Two-way ANOVA with Tukey’s post-hoc test). (J–M) Hepatic triglycerides (J), hepatic cholesterol (K), hepatic esterified cholesterol (L), and hepatic free cholesterol (M) were measured colorimetrically (n = 8–10; **p≤0.01, ***p≤0.001, ****p≤0.0001; Two-way ANOVA with Tukey’s post-hoc test). All data are presented as mean ± S.E.M., unless otherwise noted.

Figure 2—figure supplement 1. Mboat7 ASO treatment reduces Mboat7 expression in white adipose tissue (WAT), but does not dramatically alter WAT gene expression or adiposity.

C57BL/6 mice were fed chow or HFD with concurrent Control and Mboat7 ASO Injections at 12.5 mg/kg week for 20 weeks. Panels (A–K): WAT gene expression was measured using qPCR. (L) Lean and fat mass were quantified using EchoMRI imaging in HFD-fed mice.

Figure 2—figure supplement 2. Mboat7 ASO treatment does not alter Mboat7 expression in several extrahepatic tissues.

C57BL/6 mice were fed chow with concurrent Control and MBOAT7 ASO Injections at 12.5 mg/kg week for 20 weeks. Brain, spleen, heart, and skeletal muscle MBOAT7 expression was measured via qPCR (n = 5). Data are presented as mean ± S.E.M.

Figure 2—figure supplement 3. A reduction in hepatic MBOAT7 expression does not alter metabolic parameters.

C57BL/6 mice were fed chow or HFD with concurrent Control and MBOAT7 ASO injections at 12.5 mg/kg week for 12 weeks. Oxygen consumption (VO₂), carbon dioxide expiration (VCO₂), respiratory exchange ratio (RER), heat, and movement (X and Z plane) were measured using the Oxymax CLAMS indirect calorimetry system (n = 6 per group).

Figure 2—figure supplement 4. Hepatic steatosis is observed in two distinct ASOs targeting different regions of the Mboat7 mRNA.

C57BL/6 mice were fed chow or HFD with concurrent Control (non-targeting) or two distinct Mboat7 ASOs (Mboat7 ASOα or Mboat7 ASOβ) Injections at 12.5 mg/kg week for 20 weeks. Shown are representative H and E-stained liver sections, blinded pathology scoring of hepatic steatosis (n = 3 per group), and enzymatic measurement of liver triacylglycerol (TG).

Figure 2—figure supplement 5. Knockdown of Mboat7 alters plasma lipid levels in chow but not HFD-fed mice.

Male C57BL/6 mice were fed either chow or a HFD for 20 weeks while receiving concurrent ASO injections. (A and B) Plasma cholesterol (A) and triglyceride (B) levels were measured enzymatically (n = 5). (C–F) Plasma lipoprotein profile of cholesterol in chow (C, D) and HFD-fed (E, F) mice (n = 5; *p≤0.05; two-tailed t-test). Data are presented as mean ± S.E.M.