Figure 6.

SPP1 is identified as a downstream target of ALKBH5. (A) m⁶A consensus motif of GGAC is highly concentrated in SK-N-AS cells; (B) The study of m⁶A peaks showed that there were 12,508 and 10,945 in control cells and ALKBH5 knockdown, respectively, of which 1,500 were common; (C) There were 7,092 and 7,617 genes associated with the peak, with 1,931 and 2,465 uniquely associated genes, respectively; (D) Among the shared genes of differential peak-related genes of m⁶A-seq and differential genes of RNA-seq, there are 5 genes related to the ALKBH5 effect, namely SPP1, COL15A1, ADAMTS12, SYT14 and RP11-440G9.1; (E-I) When ALKBH5 was knocked down, m⁶A modification peak of the target gene was significantly increased; (J,K) RT-qPCR (J) and Western blotting (K) analysis demonstrated that SPP1 is inversely regulated by ALKBH5 in three cell types with significant differences. RT-qPCR, reverse transcription-quantitative real-time PCR. *, P <0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001.