Research Paper Volume 13, Issue 3 pp 4045—4062

ALKBH5-mediated m6A demethylation of FOXM1 mRNA promotes progression of uveal melanoma

ALKBH5 increases FOXM1 expression and FOXM1 mRNA stability in UM cells. (A) The protein levels of FOXM1 in wild-type or catalytic inactive mutation ALKBH5-expressing UM cells were measured using western blotting. (B) ALKBH5 downregulation decreased FOXM1 expression in UM cells. (C) MeRIP-qPCR analysis was used to verify ALKBH5-induced FOXM1 m6A modification. The m6A modification of FOXM1 was increased on downregulation and catalytic inactive mutation of ALKBH5 in C918 cells. (D) C918 cells were treated with dactinomycin (Act D, 2 μg/mL) to block new RNA synthesis. The stability of FOXM1 was measured by qRT-PCR at different times. Mean ± SEM, t-test, *P P P

Figure 5. ALKBH5 increases FOXM1 expression and FOXM1 mRNA stability in UM cells. (A) The protein levels of FOXM1 in wild-type or catalytic inactive mutation ALKBH5-expressing UM cells were measured using western blotting. (B) ALKBH5 downregulation decreased FOXM1 expression in UM cells. (C) MeRIP-qPCR analysis was used to verify ALKBH5-induced FOXM1 m6A modification. The m6A modification of FOXM1 was increased on downregulation and catalytic inactive mutation of ALKBH5 in C918 cells. (D) C918 cells were treated with dactinomycin (Act D, 2 μg/mL) to block new RNA synthesis. The stability of FOXM1 was measured by qRT-PCR at different times. Mean ± SEM, t-test, *P < 0.05, **P < 0.01, ***P < 0.001.