Fig. 6

METTL3 enhanced ZMYM1 mRNA expression through m6A-HuR dependent pathway. a-b RIP with anti-m6A antibody was performed in GC cells. m6A modification on ZMYM1 was depleted upon METTL3 knockdown, whereas enhanced by METTL3 overexpression. c Wild-type or mutant m6A consensus sequence was fused with firefly luciferase reporter, respectively. d The transcriptional level of wild-type ZMYM1, but not the mutation, significantly decreased in METTL3 knockdown cells. e Elevated expression of METTL3 increased the luciferase activity of wild-type ZMYM1-fused reporter, but failed to interact with mutated ZMYM1 constructs. f-g RIP-qPCR using anti-HuR antibody showed the affinity of ZMYM1 RNA to HuR in different modified METTL3 expressing cells. h-i Relative ZMYM1 and HuR mRNA levels were examined by qRT-PCR in GC cells transfected with lentiviruses carrying METTL3 and/or sh-HuR. j ZMYM1 and HuR protein levels were examined by western blot analysis in GC cells transfected with lentiviruses carrying METTL3 and/or sh-HuR. *p < 0.05, **p < 0.01