Fig. 6: Identification of P300 and SIRT1 as writers and erasers of H3K18la.

A Heatmap showing differential expression of HAT family members in proteomic data before and after DON treatment (n = 3). B CCK8 assay showing cell viability in control (CK), DON-treated, DON + Nala co-treated, and P300 knockdown or inhibition + DON + Nala co-treated porcine granulosa cells after 24 h of treatment (n = 6). C, D Western blot (WB) analysis showing the relative abundance of H3K18la after treatment with different concentrations of C646, with H3 as the loading control (n = 3). E–H WB analysis showing the effects of P300 knockdown and overexpression on H3K18la under normal conditions and after Nala supplementation. H3 and Tubulin were used as loading controls for H3K18la and P300, respectively (n = 3). I–K Confocal microscopy images showing the effects of P300 knockdown on Fe²⁺ levels, lipid peroxidation, and mitochondrial membrane potential. (scale bar:40 µm). L Quantitative data for relative fluorescence of Fe²⁺, BDP 581/591 C11, and JC-1 staining. M Schematic diagram illustrating the interaction between P300 and lactyl-CoA. Amino acids involved in the interaction are shown as stick models, the compound molecule is represented as a green stick structure, and the EP300 (A0A8W4FCE1) protein molecule is depicted as a pink helical structure. N Protein sequence alignment of P300 across different species, highlighting the high conservation of arginine at position 1068 in the HAT domain. O, P WB analysis showing the effects of the P300 R1368A mutation on H3K18la with or without 10 mM Nala. Bars indicate the mean ± SD. Data in (A and D) were analyzed using one-way ANOVA with Tukey’s post hoc test. Data in (F, H, and L) were analyzed using two-way ANOVA with Tukey’s post hoc test. Data in (P) was evaluated via an unpaired Student’s t-test. (ns, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).