Fig. 5: The m6A modification of HRAS and MEK2 mediates different mechanisms to regulate their respective protein levels. | Cell Death & Disease

Fig. 5: The m6A modification of HRAS and MEK2 mediates different mechanisms to regulate their respective protein levels.

From: Post-transcriptional modification of m6A methylase METTL3 regulates ERK-induced androgen-deprived treatment resistance prostate cancer

Fig. 5

A The results of MERIP-seq performed by C4-2 sh-con and C4-2 sh-METTL3 cell lines showed that the m6A modification sites of HRAS and MEK2. B Schematic representation of m6A position of HRAS and MEK2. C The results of MeRIP-qPCR showed that the enrichment of HRAS and MEK2 genes changed after METTL3 knockdown in C4-2/LNCap-AI cell line. D The relative ratio of protein to mRNA of HRAS and MEK2 genes in 4 cell lines. E After MG-132 and CHX were used to treat the two cell lines, respectively, the protein level of MEK2 was detected. F. Schematic representation of luciferase plasmid containing 3’UTR of HRAS, 5’UTR of MEK2 and their mutants. G After transfection of four plasmids in C4-2/LNCap-AI sh-con and C4-2/LNCap-AI sh-METTL3 cell lines for 24 h, fluorescence strength of luciferase were detected. H After four plasmids were transfected into LNCap oe-METTL3 and LNCap oe-METTL3-mut cell lines for 24 h, the fluorescence intensity of luciferase was detected. I After Act-D inhibited transcription, the RNA level of HRAS was determined after knocking down METTL3 for 2, 4 and 6 h. J RIP-qPCR was performed with IGF2BP1/2/3 antibody, and the RNA enrichment level of HRAS was detected. K. After knockdown of IGF2BP2, the RNA level of HRAS gene changed in C4-2. L After IGF2BP2 knockdown, the protein level of HRAS gene changed in C4-2. M After act-D inhibited transcription, the RNA level of HRAS decreased after IGF2BP2 knockdown, which indicated that IGF2BP2 knockdown could reduce the stability of HRAS mRNA.

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