Fig. 3: Methyltransferase activity of KMT5C is dispensable for gluconeogenesis.

a Representative western blot (left) (n = 2 per group) and densitometry analyses (right) (n = 4 per group) of indicated proteins in primary hepatocytes. Primary hepatocytes isolated from 12-week-old C57BL/6 mice were infected with lentiviruses encoding either Kmt5c or Kmt5c Mut for 48 h, followed by western blot analysis. b Glucose production using pyruvate and lactate as substrates was measured in the culture medium of primary hepatocytes treated as in (a) (n = 3 per group). c Western blot analysis (top) and protein quantifications (bottom) in primary hepatocytes (n = 3 per group). Primary hepatocytes isolated from 12-week-old female WT and KO mice were infected with Kmt5c or Kmt5c Mut lentiviruses for 48 h, followed by western blot analysis. d Western blot (left) and densitometry analyses (right) of indicated proteins in primary hepatocytes (n = 3 per group). Primary hepatocytes isolated from 12-week-old C57BL/6 male mice were treated with A-196 for 72 h. e Eight-week-old C57BL/6 male mice were injected with AAV8-TBG-Ctrl (n = 6), AAV8-TBG-KMT5C (n = 7), or AAV8-TBG-Mut KMT5C (n = 8) viruses. Blood glucose levels were measured 3 weeks after virus infusion underfeeding and 16 h fasting conditions. f, g PTT (f) (n = 6 each group, male) and GcTT (g) (AAV-Ctrl, n = 6; AAV-KMT5C, n = 7; AAV-Mut, n = 7, male) were performed in mice of (e). h, i Quantitative PCR (h) (AAV-Ctrl, n = 6; AAV-KMT5C, n = 7; AAV-Mut, n = 8, male) and western blot (i) (AAV-Ctrl, n = 5; AAV-KMT5C, n = 5; AAV-Mut, n = 4, male) detected gluconeogenic gene expression in mice of (e). The densitometry analysis was shown in (i, right). Mice were fasted for 16 h before the assay. j, k Representative KMT5C (j) (WT + IgG, n = 2; WT + αKMT5C, n = 5; KO + IgG, n = 2; KO + αKMT5C, n = 5, male) and H4K20me3 (k) (WT + IgG, n = 2; WT + αH4K20me3, n = 4; KO + IgG, n = 2; KO + αH4K20me3, n = 5, male) ChIP assays performed in the WT and KO mice livers after 24 h fasting. Zfp560 and Rdna loci were used as positive controls for H4K20me3 enrichment. a–k data present mean ± s.e.m. a–d n represents biologically independent cell samples; e–k n presents biologically independent animals. a–k statistical analysis was performed using one-way ANOVA. All experiments here were repeated independently three times with similar results. Source data are provided as a Source Data file.