Fig. 4: Intracellular lactate levels were increased by glycolytic process after Fam172a knockdown.
From: Histone lactylation mediated by Fam172a in POMC neurons regulates energy balance
A, B Neuro2a cells were transfected with ADV-shCtrl or ADV-shFam172a at 37°Cfor 48 h, and then RNA-Seq was performed. Heatmaps of the top 15 genes with significant differences (A). Top ten KEGG pathways were also shown (B). n = 3 cell cultures per group. C Schematic of the glycolytic pathway, and highlighting (red) the participating enzymes. D, E Neuro2a cells were transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h, total RNAs or proteins were then extracted, and the relative mRNA (D) and protein (E) levels of the indicated genes were assessed. n = 5 cell cultures per group for qPCR or 3 cell cultures per group for western blot. F, G Seahorse metabolic analysis (ECAR) of neuro2a cells which were transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h. n = 5 cell cultures per group. H Intracellular lactate levels of neuro2a cells transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h. n = 4 cell cultures per group. I, J Infection with ADV-shFam172a increased mouse LDHA (I) and PDK1 (J) promoter activity in Neuro2a cells. n = 5 cell cultures per group for LDHA promoter activity analysis or 6 cell cultures per group for PDK1 promoter activity analysis. K, L Neuro2a cells were transfected with CMV-Ctrl or CMV-Fam172a at 37°C for 48 h, total RNAs or proteins were then extracted, and the relative mRNA (K) and protein (L) levels of the indicated genes were assessed. n = 5 cell cultures per group for qPCR or 3 cell cultures per group for western blot. M, N Seahorse metabolic analysis (ECAR) of neuro2a cells transfected with CMV-Ctrl or CMV-Fam172a at 37°C for 48 h. n = 5 cell cultures per group. O Intracellular lactate levels of neuro2a cells which were transfected with CMV-Ctrl or CMV-Fam172a at 37°C for 48 h. n = 5 (CMV-Ctrl) or 4 (CMV-Fam172a) cell cultures per group. Data are presented as mean ± SEM. Hypergeometric test with Bonferroni’s post hoc test (B), two-tailed Student’s t-test (D, G, H, I, J, K, N, O). Source data are provided as a Source Data file.
