Fig. 3: FTO mediates the stability of PHF1 mRNA by demethylating m6A process. | Cell Death Discovery

Fig. 3: FTO mediates the stability of PHF1 mRNA by demethylating m6A process.

From: Down-regulated m6A reader FTO destabilizes PHF1 that triggers enhanced stemness capacity and tumor progression in lung adenocarcinoma

Fig. 3

A MeRIP assay was used to detect the m6A abundance on PHF1 mRNA in A549 and PC-9 cells. B Gene correlation analysis of PHF1 and m6A-related enzymes using Spearman statistics in TCGA-LUAD cohort. C Western blotting assay showing the protein levels and associations of FTO and PHF1 in LUAD cells. D RIP assay showing the endogenous interaction between FTO and PHF1 pre-mRNA using the nuclear extracts of A549 cells expressing FTO with or without Flag tag. E MeRIP-qPCR analysis indicating the m6A modification of PHF1 pre-mRNA in FTO-KD cells with or without WT or R316Q mutant expression. F FTO-KD A549 cells with or without WT or R316Q mutant expression were treated by actinomycin D at the different timepoints. The levels of remaining PHF1 mRNA were determined via qPCR assay. G RIP assay showing the interaction between endogenous YTHDF2 and PHF1 pre-mRNA in A549 cells with or without FTO-KD. H RIP analysis of transcripts from nuclear extracts of control and FTO-KD A549 cells expressing YTHDF2 or Flag-YTHDF2. I The qPCR assays detecting the PHF1 mRNA levels in FTO-KD A549 and PC-9 cells treated with or without YTHDF2 siRNA. J The PHF1 3′UTR firefly luciferase activity was detected in FTO-KD A549 and PC-9 cells treated with or without YTHDF2 siRNA. K FTO-KD A549 cells treated with or without siYTHDF2 were cultured with actinomycin D at the indicated timepoints. The qPCR assays were performed to detect the amount of remaining PHF1 levels after the above treatment. Bar = Mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

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