Figure 4.

The Expression Level of AK4 Protein Is Modulated by the m⁶A Writer Protein METTL3

(A and B) Images from western blot for monitoring the changes in AK4 protein after overexpression of METTL3 in MCF-7 cells (A), or knockdown of METTL3 in TamR MCF-7 cells (B). (C) Western blot for examining the changes in expression of AK4 protein after overexpression of YTHDF2 or YTHDF2 W432A in TamR MCF-7 cells. (D–F) Quantification of AK4 protein change based on results obtained from (A) (A)–(C), respectively. The values in the bar graphs represent the values that were normalized against the band intensities of GAPDH. (G and H) Parental MCF-7 cells with stable overexpression of METTL3 (G) and TamR MCF-7 cells with stable knockdown of METTL3 (H) were treated with serially diluted 4-OHT for 72 h and the cell viability was measured using CCK8. The data were normalized against the control groups (ethanol). The data in (D)–(H) represent mean ± SD (n = 3). The p values were calculated by using the unpaired, two-tailed Student’s t test. ^#p ≥ 0.05, ∗0.01 ≤ p < 0.05, ∗∗0.001 ≤ p < 0.01, ∗∗∗p < 0.001.