Fig. 5

MA2 downregulated CDK2 expression and cell viability through the 3’UTR-m⁶A mediated mRNA degradation. a Schematic diagram of plasmids used in the 3’UTR fluorescence reporter assay. Letter A and T (red) represented the mutation of predicted m⁶A to T. b Effects of CDK2-3’UTR and CDK2-mut on the expression of mCherry. Cells were treated with 40 and 80 μmol/L of MA2, and simultaneously transfected with CDK2-3’UTR and CDK2-mut plasmids. After 48 h, the cells were photographed under the fluorescence microscope. Bar = 50 μm. c Relative fluorescence intensity was showed in column diagram. Data were represented by the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. d Relative mRNA level of mCherry to GFP in each group. n = 3 independent duplications, *P < 0.05, **P < 0.01. e Cells were treated with 0, 40 and 80 μmol/L MA2 for 24 h. After MA2 treatment, cells were treated with 5 μmol/L actinomycin D for 4 h and 8 h, respectively. Remained RNA level was quantified by RT-PCR. Data were represented by the mean ± SEM, n = 3. *P < 0.05, **P < 0.01