Fig. 2.

OGT^LKOmice exhibit liver extensive fibrosis at 8 weeks.

Male liver-specific Ogt knockout mice (OGT^LKO) and control floxed littermates (OGT^LWT) were studied 8 weeks after birth in the fed state. (A) Fed blood glucose (mmol/L), body weight (g), liver weight, and spleen weight are shown. Liver and spleen weights are represented as percentage of body weight. (B) Macroscopic view of spleen and liver from OGT^LWT (left) and OGT^LKO (right) mice. (C) Macroscopic view of histological liver section stained with Sirius Red. (D) Relative expression levels of fibrosis markers normalised to TBP. (E) Expression levels of OGT normalised to TBP. (F) Western blots of OGT content and O-GlcNAcylation levels in livers of OGT^LWT and OGT^LKO mice at 8 weeks. Three representative samples are shown. Actin was used for normalisation. (G) Liver sections stained with H&E, Sirius Red, ɑSMA, OGT, Ki67, and TUNEL. Scale bars = 100 μm. (H) Western blot analysis of necroptosis proteins (MLKL and RIPK3). Six representative samples are shown. Tubulin was used for normalisation and quantification. (I) Serum levels of ALT and LDH. (J) Relative expression levels of inflammatory markers normalised to TBP. (K) Serum levels of inflammatory cytokines. Data are shown as mean ± SEM of 13–15 male mice per condition. ∗p <0.05, ∗∗p <0.01, ∗∗∗p <0.001 by unpaired Student’s t test (Mann–Whitney U test). ɑSMA, alpha-smooth muscle actin; ALT, alanine aminotransferase; Ccl5, C–C motif chemokine ligand 5; Col3a1, collagen type III alpha 1 chain; Col6a1, collagen type VI alpha 1 chain; LDH, lactate dehydrogenase; Mcp1, monocyte chemoattractant protein-1; MLKL, mixed lineage kinase domain-like pseudokinase; OGT, O-GlcNAc transferase; RIPK3, receptor-interacting serine/threonine-protein kinase 3; TBP, TATA-box binding protein; Tgfβ, transforming growth factor beta; Tnfɑ, tumour necrosis factor alpha; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labelling.