N6-methyladenosine reader IGF2BP2 regulates NIPSNAP1-mediated mitophagy and mitochondrial dynamics to alleviate hepatic ischemia-reperfusion injury

Figure 4 Insulin-like growth factor 2 mRNA-binding protein 2 regulates 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1 mRNA stability through n6-methyladenosine modification. ^aP < 0.05, ^bP < 0.01. A: The n6-methyladenosine enrichment of 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1 (NIPSNAP1) mRNA in HepG2 cells was assessed by MeRIP^TM-quantitative real-time polymerase chain reaction, n = 6; B: Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) was pulled down and enriched with biotinylated NIPSNAP1 mRNA in HepG2 cells; C: IGF2BP2 RNA immunoprecipitation was used to assess the interaction between IGF2BP2 and NIPSNAP1 mRNA in HepG2 cells via quantitative real-time polymerase chain reaction, n = 6; D and E: HepG2 cells were subjected to hypoxia/reoxygenation after transfection with IGF2BP2 small interfering RNAs. NIPSNAP1 mRNA levels, n = 6. IGF2BP2 and NIPSNAP1 proteins, n = 3; F and G: HepG2 cells were subjected to hypoxia/reoxygenation after transfection with pcDNA-IGF2BP2. NIPSNAP1 mRNA levels, n = 6. IGF2BP2 and NIPSNAP1 proteins, n = 3; H: After HepG2 cells were transfected with si-control or si-IGF2BP2, they were treated with 2 μg/mL actinomycin D, and total RNA was extracted at different time intervals. NIPSNAP1: 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1; IGF2BP2: Insulin-like growth factor 2 mRNA-binding protein 2; IB: Immunoblotting; NC: Negative control; H/R: Hypoxia/reoxygenation.