Figure 4:

Impact of high fat diet on hepatic lipid content of Tm6 LKO mice and rescue with AAV8 Tm6sf2. A. Hepatic triglyceride content in male and female Tm6 Flox and Tm6 LKO mice fed HMFD for 3 weeks (female, n=6/genotype; male, n= 11–15). A similar phenotype was found using Tm6 LKO line 2. B. AAV-mediated expression of Flag Tm6sf2. Left: Relative expression of Tm6 mRNA in livers of Tm6 LKO mice (n=6–8/group) transduced with LacZ AAV, WT Tm6 AAV, or E167K AAV, normalized to expression of endogenous Tm6sf2 in Tm6 Flox livers. All groups are significantly different compared to flox controls. Right: Western blot showing expression of Flag tagged Tm6 in AAV transduced liver tissue using anti-Flag antibody. Expression of Gapdh protein is shown as a loading control. C. Left: Hepatic TG content in liver of HMFD- fed Tm6 Flox and Tm6 LKO mice transduced with AAV constructs (n=6–10 males/group). Right: Quantitation of lipid droplet size in Tm6 LKO mice transduced with Lac Z or WT Tm6 AAV. D. VLDL secretion in HMFD- fed male and female Tm6 Flox and Tm6 LKO mice transduced with Lac Z, Tm6 WT, and E167K AAV (n=3–4/group). E. Expression of lipogenic genes in Tm6 Flox and AAV-transduced LKO mice fed HMFD (n=5–7 animals/group). F. Sirius red stained fibrotic area in Tm6 Flox and Tm6 LKO mice transduced with AAV (n=5–6/group). Representative images are shown (400x magnification, scale bar = 20μM). For all panels, asterisks indicate p<0.05.