Fig. 7.

KD improves oxidative stress, ER stress, DNA damage, hepatic fibrosis, and liver injury in OGT^LKOmice.

Liver-specific Ogt knockout mice (OGT^LKO) and control floxed littermates (OGT^LWT) (males and females) were weaned (at 3 weeks of age) on KD and sacrificed 5 weeks later in the fed state. (A) Relative expression levels of oxidative stress markers normalised to TBP. (B) The proportion of highly polyploid (≥8n) mononuclear hepatocytes relative to total hepatocytes was quantified in each group. Blue: plasma membrane labelling (KL1). Red: nuclear labelling (Hoechst) (scale bar = 100 μm). (C) Western blot analysis of CHOP protein content in livers of OGT^LWT and OGT^LKO mice. Three to four representative samples are shown. (D) Western blot analysis of γH2AX protein content in livers of OGT^LWT and OGT^LKO mice. Three to four representative samples are shown. (E) Quantification of CHOP and γHA2X are shown. Actin was used as a loading control. (F) Liver sections stained with H&E, F4/80, SOX9, Krt7, Sirius Red, and OGT. Scale bars = 100 μm. (G) Quantification of fibrosis by SHG microscopy. (H) Relative expression levels of fibrotic markers normalised to TBP. (I) Relative expression levels of inflammatory markers normalised to TBP. (J) Serum levels of ALT and ALP. Data are shown as mean ± SEM of four to eight mice (males and females). ∗p <0.05, ∗∗p <0.01, ∗∗∗p <0.001, ∗∗∗∗p <0.0005 by two-way Anova followed by Bonferroni post hoc test._. ɑSMA, alpha-smooth muscle actin; ALP, alkaline phosphatase; ALT, alanine aminotransferase; CHOP, CCAAT–enhancer-binding protein homologous protein; Col3a1, collagen type III alpha 1 chain; Col6a1, collagen type VI alpha 1 chain; ER, endoplasmic reticulum; Gsta1, glutathione S-transferase alpha 1; Gstm3, glutathione S-transferase mu 3; H2AX, H2AX variant histone; KD, ketogenic diet; Krt7, cytokeratin 7; Mcp1, monocyte chemoattractant protein-1; NqO1, NAD(P)H quinone dehydrogenase 1; Nrf2, nuclear factor erythroid-derived 2-related factor; OGT, O-GlcNAc transferase; SD, standard diet; SHG, second harmonic generation; SOX9, sex-determining region Y-box 2; TBP, TATA-box binding protein; Tgfβ, transforming growth factor beta; Tnfɑ, tumour necrosis factor alpha.