Fig. 3

Increased P300 mediates the high expression of METTL1 via H3K27 acetylation modification. A Data from the UCSC genome bioinformatics site indicated high H3K27ac enrichment in the promoter of METTL1. B CHIP-qPCR was used to confirm the difference in the H3K17ac-modified METTL1 promoter region between HSPC and CRPC cells. C-D The mRNA levels of METTL1 in C646 (20 µM)-treated LNCaP-AI (C), and C4-2 (D) cells at the indicated time points were measured by RT-qPCR. E-F The mRNA levels of METTL1 in different concentrations of C646 (48 h)-treated LNCaP-AI (E), and C4-2 (F) cells were measured by RT-qPCR. G The METTL1 and H3K27ac protein levels were measured via western blot assay after C646 treatment (20 µM) for 48 h. H IHC was carried out to detect the P300 level in TMA containing HSPC and CRPC patients’ samples. I Pearson correlation between METTL1 and P300 (R = 0.58). J-K The mRNA change of METTL1 after P300 was inhibited using small interfering RNA (siRNA). L The P300, METTL1, and H3K27ac protein levels were measured via western blot assay after P300 suppression. M-N CHIP-qPCR was used to determine the enrichment of H3K27ac at the promoter of METTL1 in control or P300 deficiency LNCaP-AI and C4-2 cells. *P < 0.05; **P < 0.01; ***P < 0.001. H3K27ac, H3K27 acetylation; CHIP, chromatin immunoprecipitation