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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

Fig. 4

RBM15 participated in the methylation of TMBIM6 in the form of m6A modification. a The methylated RNA (m6A) level was determined after RBM15 knockdown in the LSCC cell lines. b Hierarchical clustering for mRNAs with differential “m6A quantity”. c Coalition analysis of significant changes in RNA expression level and m6A level in pairs of LSCC specimens. d The expression of selected mRNAs was investigated by qRT-PCR in AMC-HN-8 cells transfected with shCtrl or shRBM15. e The expression of selected mRNAs was investigated by qRT-PCR in TU-212 cells transfected with vector or RBM15-expressing plasmid. f MeRIP assay was performed to identify whether m6A modification was enriched in the TMBIM6 sequence. g and h MeRIP assays were performed to identify variation in m6A modification enrichment in TMBIM6 after silencing or overexpressing RBM15 in LSCC cells. i and j qRT-PCR analysis of TMBIM6 mRNA after RBM15 inhibition or overexpression. k Diagram summarizing the mechanism of MazF RNA endonuclease. MazF cleaves only nonmethylated RNA, sites with m6A methylation remained uncleaved. l A schematic diagram showing the methylation site on TMBIM6 and mutates this site. m The luciferase activities in AMC-HN-8 cells co-transfected with TMBIM6-WT or TMBIM6-Mut together with shCtrl or shRBM15. n The luciferase activities in TU-212 cells co-transfected with TMBIM6-WT or TMBIM6-Mut together with Vector or RBM15. o and p qRT-PCR verified the expression of TMBIM6 in 34 pairs of LSCC tissues and cell lines. q Correlation analysis was conducted on the expression levels of RBM15 and TMBIM6 in 34 cases of LSCC tissues. r Correlation analysis was performed on the expression levels of RBM15 and TMBIM6 in the TCGA database. s The analysis results of the TCGA database showed the relationship between the expression level of TMBIM6 in HNSC patients and the overall survival rate. t The expression efficiency of TMBIM6 after transfection with shCtrl or shTMBIM6 in LSCC cells. u The expression efficiency of TMBIM6 after transfection with Vector or TMBIM6 in LSCC cells

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