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Fig. 4 | Molecular Cancer

Fig. 4

From: The m6A demethylase ALKBH5 promotes tumor progression by inhibiting RIG-I expression and interferon alpha production through the IKKε/TBK1/IRF3 pathway in head and neck squamous cell carcinoma

Fig. 4

ALKBH5 regulates RIG-I expression through m6A modification. a The top 15 biological processes are shown with a bubble chart. b, c Candidate differentially expressed genes were detected in Cal27 and HN4 cells after ALKBH5 silencing using qRT-PCR. d The m6A peak visualization of m6A-seq in DDX58 transcripts in Cal27 cells with or without ALKBH5 depletion is shown. The m6A peaks are in the 3’UTR of DDX58. e Methylated RNA in cells with or without ALKBH5 depletion was immunoprecipitated with an m6A antibody, followed by qPCR analyses with primers against DDX58 mRNA. (f) RIG-I, encoded by DDX58, was detected by immunoblotting assays after ALKBH5 silencing for 48 h. g RIG-I expression was detected by immunoblotting assays after siRNA cotransfection for 48 h. h RIG-I expression was detected by immunoblotting assays after ALKBH5 or H204 mutant transfection for 48 h. i RIG-I protein expression was detected after plasmid cotransfection for 48 h. j Luciferase vectors with the wild-type (WT) or mutated m6A nucleotides (MT) in the DDX58 gene were transfected into 293 T cells after ALKBH5 depletion. Relative luciferase activity was measured. k DDX58 mRNA expression was detected after incubation with 5 μg/mL actinomycin D for the indicated times and normalized to GAPDH. l The mRNA half-lives were estimated according to linear regression analysis after the indicated actinomycin D treatment. Data information: Data are presented as the mean ± S.D. One-way ANOVA; *P < 0.05, **P < 0.01

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