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Fig. 2 | Molecular Cancer

Fig. 2

From: m6A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2–mediated YAP activity in NSCLC

Fig. 2

ALKBH5 controls YAP expression by regulation m6A level in NSCLC. (a) Pie chart depicting the fraction of m6A peaks in 4 transcript segments. (b) Gene ontology (GO) enrichment analysis of tumor methylated genes in NSCLC patients. (c) The m6A levels of total and YAP were detected in human NSCLC cell lines determined by m6A methylation quantification kit and MeRIP-qPCR assay. (d) The total m6A levels were higher in 30 paired fresh NSCLC tumor cancer tissues (Tumor) than matched adjacent normal tissues (Normal) (n = 30). (e) The interaction between ALKBH5 and YAP pre-mRNA was detected by RIP. (f) The relative m6A level of YAP pre-mRNA was detected by MeRIP-qPCR in A549 cells. (g) Sequence motifs in m6A peaks identified by using m6Avar database from YAP CDS. (h) Putative m6A modification sites in the CDS sequence of YAP and synonymous mutations in the YAP CDS. (i) The interaction between ALKBH5 and YAP pre-mRNA was detected by RIP. (j) The relative of m6A level in YAP from co-expression of ALKBH5 and YAP WT/Muts# (left panel) or ALKBH5 and YAP Muts (right panel) in A549 and H1299 cells. (k) The relative mRNA levels of YAP were analyzed by RT-PCR. (l) The mRNA and protein levels of ALKBH5, YAP, CTGF and Cyr61 were detected in A549 cells. (m) The negative correlation between ALKBH5 and YAP was analyzed from TCGA database. (n) The expressions of wild type (ALKBH5 WT) and catalytic mutant (ALKBH5 KD) of ALKBH5 were analyzed by RT-PCR and western blot assays. (o) The relative of m6A level in YAP from ALKBH5 WT or KD transfected A549 cells. (p) The mRNA levels of ALKBH5, YAP, CTGF and Cyr61 were detected in A549 and H1299 cells. Results were presented as mean ± SD of three independent experiments. *P < 0.05 or **P < 0.01 indicates a significant difference between the indicated groups. ns, not significant

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