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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Biochem Pharmacol. 2018 Dec 19;160:134–145. doi: 10.1016/j.bcp.2018.12.014

Fig. 4. m6A methylation in the mRNA of p53 R273H.

Fig. 4.

A, Silencing METTL3 enhanced pp53 levels in TP53-Dox cells. TP53-Dox cells were treated with siRNA against METTL3 (100 nM, 6 days) and PDMP (5 μM, 6 days). Equal amounts of detergent-soluble proteins extracted (50 μg/lane) were resolved using 4–20% gradient SDS-PAGE, and then immunoblotted with corresponding antibodies. pp53, phosphorylated p53 (Ser15); METTL3, methyltransferase-like 3. Protein levels are represented below bands, as their OD values normalized against GAPDH from three settings of blots and compared to vehicle treatment. *, p<0.001 compared to vehicle. B, Methylation to m6A catalyzed by METTL3. C, RNA m6A methylation in cells. Equal amounts of total RNA extracted from cells of SW48-Dox (wt p53), TP53-Dox (TP53; R273H/+), WiDr (R273H+/+) and TP53-Dox treated with neplanocin A (NPC, 20 nM, 6 days) or PDMP (5 μM, 6 days) were assessed with MeRIP RT-qPCR. *, p<0.001, compared to SW48-Dox cells; **, p<0.001, compared to vehicle control in TP53-Dox cells. D, Real-time PCR chart, as described above.