Fig. 5
From: Demethylase ALKBH5 suppresses invasion of gastric cancer via PKMYT1 m6A modification
ALKBH5 and IGF2BP3 together regulates the expression of PKMYT1 via its m6A modification (A) Results of mass spectrometry using PKMYT1 probe. (B) RNA pulldown of endogenous IGF2BP3 using NC or PKMYT1 probe. (C) RIP-qPCR assay of PKMYT1 enrichment by IGF2BP3. (D-E) Representative IHC pictures of IGF2BP3 in GC tissue microarray (TMA) and the comparison of area density in IGF2BP3-staining (scale bars = 100 µm). (F) MRNA expression comparison of IGF2BP3 in TCGA database. (G) Kaplan–Meier OS analysis of IGF2BP3 expression in patients with GC (HR = 1.23, p = 0.021, log-rank test) (H) Correlation analysis of PKMYT1 and IGF2BP3 expression in TMA. Pearson r = 0.2151, p < 0.0001. (I) Expression correlation between PKMYT1 and IGF2BP3 in STAD of TCGA database. Pearson r = 0.17, p = 0.00024. (J-K) The mRNA and protein level of PKMYT1 in si-IGF2BP3 HGC-27 cell. (L-N) RNA stability of PKMYT1 mRNA in ALKBH5-overexpressing, ALKBH5 knockout and H204A GC cells after treated with actinomycin D (5 µg/mL). (O) MRNA level of PKMYT1 at the indicated time points after actinomycin D treatment in si-IGF2BP3 HGC-27 cell. (P) RNA stability of PKMYT1 mRNA in PKMYT1-CDS, CDS-mut1 and CDS-mut2 groups. (Q-R) RNA-pulldown and RIP assay between PKMYT1 mRNA and IGF2BP3 after mutation in PKMYT1-CDS