Research Paper Volume 15, Issue 18 pp 9590—9613
YY1 is regulated by ALKBH5-mediated m6A modification and promotes autophagy and cancer progression through targeting ATG4B
Figure 4. ALKBH5 weakens the m6A levels of YY1 mRNA. (A) MeRIP assay showing the YY1 m6A levels in human GC cell lines compared to GES-1 cells. (B, C) qRT-PCR and MeRIP assays displaying the mRNA and m6A levels in MGC803 and AGS cells induced for the overexpression or knockdown of ALKBH5. (D) Schematic illustration showing the m6A modification site (AAGGGA, AAAAGA) at position of 130 and 290 base on the YY1 3’-UTR from SRAMP Browser, and the wild-type (WT) or mutation (Mut) of YY1 3’-UTR reporters were designed (m6A was replaced by T). (E, F) Dual-luciferase and Western blot assays revealing the 3’-UTR activity and protein levels of YY1 in GC cells stably transfected with Ca-NC, Ca-ALKBH5, Ci-NC or Ci-ALKBH5. (G, H) Dual-luciferase and qRT-PCR assays showing the binding activity and mRNA half-life of YY1 in GC cells stably transfected with Ca-NC, Ca-ALKBH5, Ci-NC or Ci-ALKBH5. *P < 0.05 vs. GES-1, Ca-NC, Ci-NC.