Distinct N6-methyladenosine (m6A) Regulation of Innate Immune Responses During Bacterial Infections II

N6-methyladenosine (m6A) is a dynamic post-transcriptional RNA modification influencing all aspects of mRNA biology. Here, we examined cellular m6A epitranscriptomes during P.aeruginosa infection to identify m6A-regulated innate immune response genes with or without ALKBH5 knockdown. We showed that a significant portion of cellular genes including many innate immune response genes underwent m6A modifications in 5’UTR and 3’UTR, from which we identified common and distinct m6A-modified genes under different stimulating conditions. RAW264.7 cells at 4×10^5 cells per mL were infected with P. aeruginosa at 107 per mL and harvested at 0, 2, 4, and 6 h post treatment. RAW264.7 cells with ALKBH5 knockdown were infected with P. aeruginosa 6 hr post infection. MeRIP was performed to enrich m6A methylated mRNA with m6A antibody, and sequenced with corresponding inut RNA.