Fig. 5: METTL1 regulates ATF3 expression mediated by the process of miR-760. | Cell Death Discovery

Fig. 5: METTL1 regulates ATF3 expression mediated by the process of miR-760.

From: METTL1 drives tumor progression of bladder cancer via degrading ATF3 mRNA in an m7G-modified miR-760-dependent manner

Fig. 5

A Volcano plot showed differentially expressed mRNAs upon METTL1 depletion from mRNA-seq. (|log2FC| > 1, P value < 0.05) 800 mRNAs upregulated (red) and 1177 mRNAs downregulated (blue). B Heatmap showed a part of upregulated mRNAs from mRNA-seq. C KEGG enrichment analysis of differentially expressed mRNAs upon METTL1 depletion from mRNA-seq. D Venn diagram showed the overlap among upregulated mRNAs upon METTL1 depletion from mRNA-seq, candidate genes of miR-760 in miRNA online databases and genes negatively correlated with METTL1 in TCGA database. E RT-qPCR assay confirmed the expression of ATF3 was negatively correlated with the expression of METTL1. GAPDH was used for the normalization control. The average of three independent biological replicates ±SDs is shown (*P < 0.05, ***P < 0.001). F Pearson correlation analysis showed the expression of ATF3 was negatively correlated with the expression of METTL1 in TCGA database. (r = −0.2049, P value < 0.0001). G Western blot assay showed the elevated protein expression of ATF3 upon METTL1 depletion. GAPDH was used for the normalization control. A representative experiment of three independent biological replicates is shown. H RT-qPCR assay showing ATF3 was downregulated by overexpression of miR-760. The average of three independent biological replicates ±SDs is shown (*P < 0.05, ***P < 0.001). I Western blot assay showing ATF3 was downregulated by overexpression of miR-760 at protein expression level. GAPDH was used for the normalization control. A representative experiment of three independent biological replicates is shown. J Schematic showing the target site of miR-760 in 3’UTR of ATF3 mRNA was muted as presented. K Dual-luciferase reporter assay indicated miR-760 significantly suppressed the luciferase activity of vectors carried 3’UTR of ATF3. The average of three independent biological replicates ±SDs is shown (***P < 0.001). L Dual-luciferase reporter assay indicated depleted METTL1 enhanced luciferase activity of vectors carried 3’UTR of ATF3, which was targeted by miR-760 in UM-UC3. The average of three independent biological replicates ±SDs is shown (**P < 0.01, ***P < 0.001). M Western blot assay showing the rescue of ATF3 upregulation upon transfection with miR-760 in METTL1-depleted cells. A representative experiment of three independent biological replicates is shown.

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