Fig. 5

YTHDF2 recruits DDX6 to promote NR4A1 mRNA decay. A, B KEGG and GO pathway enrichment analysis of the high-confidence interacting proteins with YTHDF2. C Overlap of RNA degradation related genes and cytoplasmic mRNA processing body related genes as identified by A and B. D The interaction of YTHDF2 protein and DDX6 protein via Co-IP assay. E Confocal microscopy images indicated that YTHDF2 (red) were colocalized with P-bodies marker DDX6 (green). F-G, The NR4A1 mRNA (F) and protein (G) were determined by RT-qPCR and western blot assays upon silencing DDX6 in HeLa cells. H The mRNA stability of NR4A1 mRNA in HeLa cells after treatment with act-D during indicated time points in shR-DDX6 and shR-NC HeLa cells. I RIP-qPCR was used to determine the enrichment of NR4A1 in shR-NC and shR-YTHDF2 HeLa cells by using a DDX6-antibody. J The effect of YTHDF2 on DDX6 protein expression via western blot assay. K, RIP-qPCR was performed to measure the enrichment of NR4A1 in DDX6 knockdown and control cells using a YTHDF2-antibody. All experiments were performed at least 3 independent times, and data are presented as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001