Fig. 7

YTHDF2-NR4A1 axis meditates the transcription repression of AKT1 by recruiting LHC complex. A YTHDF2 was positively correlated with AKT1 in GEPIA2 database. B The mRNA levels of AKT1 were detected by RT-qPCR with overexpression and knockdown of YTHDF2. C Western blot assay was used to detect the alteration of AKT and AKT phosphorylation after overexpression and knockdown of YTHDF2 in CC cells. D RIP assay of the enrichment of AKT1 transcripts by YTHDF2. E RNA remaining for AKT1 in HeLa cells transfected with control and YTHDF2 knockdown were determined by RT-qPCR after treated with act-D at indicated times. F GSEA identified a significant association between NR4A1 and AKT pathway. G Western blot showed the protein levels of AKT1 and p-AKT1 after overexpression and knockdown of NR4A1 in CC cells. H Co-IP assay was performed to analyze the interaction between NR4A1 protein and SP1 protein. I–K The analysis of SP1-bound site of AKT1 promoter. L The EGFP reporter assay was preformed to measure the EGFP levels by co-transfected with the indicated plasmids. β-actin (left) and RFP (right) were used for normalization. M Binding of NR4A1 to LSD1, CoREST and HDAC1 in HeLa cells as shown by Co-IP assay. N ChIP assay of AKT1 promoter binds of NR4A1 and components of LHC complex. O The mRNA and protein expression were measured by RT-qPCR (left) and western blot assay (right) after transfected of shR-LSD1, shR-CoREST and shR-HDAC1, respectively, upon a NR4A1-overexpression background. P All findings in this study are presented as a schematic diagram. 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