Fig. 7: Downregulation of H3K18la suppresses iron homeostasis and ovarian steroidogenesis-related gene expression.

A Schematic diagram of the CUT&Tag workflow in DON-treated granulosa cells. B, C Genome-wide distribution of H3K18la and its enrichment near transcription start sites (TSS). Schematic of the experimental design illustrating the comparison between control (CK) and DON. Three biological replicates were performed for each group, and CUT&Tag sequencing was conducted to analyze H3K18 lactylation binding at gene promoters. D Heatmap showing differential H3K18 lactylation binding peaks at gene promoters, as assessed by CUT&Tag. The heatmap illustrates changes in lactylation modifications between CK and DON groups. E Volcano plot highlighting the differentially expressed genes (DEGs) between CK and DON conditions, with genes categorized based on their lactylation status (gain, loss, stable). The plot shows significant changes in gene expression associated with H3K18 lactylation following sleep deprivation. F KEGG analysis of genes associated with promoter-enriched LOSS DPs. G, H IGV visualization showing the enrichment of H3K18la at the promoter regions of STEAP3 and HSD3B1. The tracks show clear changes in lactylation patterns at these gene promoters. I Enrichment ratio of H3K18la at the promoter regions of STEAP3 and HSD3B1 in DON-treated and DON + Nala co-treated granulosa cells (n = 3). J–L Effects of DON treatment and DON + Nala co-treatment on the gene and protein expression levels of STEAP3 and HSD3B1. (Scale bar:40 µm) Bars indicate the mean ± SD. Data were analyzed using a two-way ANOVA with Tukey’s post hoc test, with significance levels denoted as follows: ns, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (A) was created with FigDraw with copyright.