Fig. 6: Lactate regulates the transcriptional level of PAM through H4K12la.

A Western blot analysis of H4K12la in neuro2a cells treated with NaCl or lactate at 37°C for 24 h. n = 3 cell cultures per group. B The binding density of H4K12la was visualized by deepTools: the heatmap presents the CUT&Tag tag counts on the different H4K12la binding peaks in neuro2a cells treated with NaCl or lactate at 37°C for 24 h, ordered by signal strength. C Genome-wide distribution of upregulated H4K12la-binding peaks in neuro2a cells treated with NaCl or lactate at 37°C for 24 h. D Genome browser tracks of CUT&Tag signals at the PAM target gene loci. E Neuro2a cells were treated with NaCl or latate at 37°C for 24 h, total RNA was then extracted, and the relative mRNA levels of the indicated genes were assessed. n = 6 (Ctrl) or 5 (Lactate) cell cultures per group. F Lactate treated neuro2a cells presented increased mouse PAM promoter activity. n = 5 (Ctrl) or 4 (Lactate) cell cultures per group. G A proposed working model for how lactate influenced the transcription of PAM. Created in BioRender. Chen, Z. (2024) https://BioRender.com/j97w406. H Neuro2a cells were transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h, after which RNA-Seq was performed. Heatmaps of POMC to α-MSH pathway target genes. n = 3 cell cultures per group. I Neuro2a cells were transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h, total RNA was then extracted, and the relative mRNA levels of the indicated genes were assessed. n = 6 (ADV-shCtrl) or 5 (ADV-shFam172a) cell cultures per group. J Knockdown of Fam172a in neuro2a cells increased mouse PAM promoter activity. n = 5 cell cultures per group. K Western blot analysis of PAM and H4k12la in neuro2a cells transfected with ADV-shCtrl or ADV-shFam172a at 37°C for 48 h. n = 3 cell cultures per group. Data are presented as mean ± SEM. two-tailed Student’s t-test (E, F, I, J). Source data are provided as a Source Data file.