Figure 6.

SPI1 inhibited the transcriptional activity of FTO

(A) Predicted SPI1 DNA-binding sequences in the FTO promoter region. (B) The correlation between FTO and SPI1 and the correlation between SPI1 and MTMR3 in TCGA and CGGA databases. Statistical analysis was determined using Pearson correlation coefficient analyses. (C) The relative mRNA expression of SPI1 (left) and FTO (right) in GBM cells transfected with siNC, siSPI1, or siSPI1-2. (D) The protein expression of SPI1, FTO, MTMR3, N-cad, CD44, PCNA, and vimentin in GBM cells transfected with siNC, siSPI1, or siSPI1-2. (E) ChIP-PCR of the FTO promoter region in GBM cells. (F) Relative FTO promoter luciferase activity in GBM cells transfected with siNC, siSPI1-1, or siSPI1-2. (G) Relative FTO promoter luciferase activity in the indicated GBM cells with or without the mutation of the SPI1 binding motif. (H) Transwell assays of U87MG and U118MG GBM cells transfected with siNC or siSPI1 in the presence or absence of FTO as indicated. Scale bar, 50 μm. (I) CCK-8 assays of U87MG and U118MG GBM cells transfected with siNC or siSPI1 in the presence or absence of FTO as indicated. Comparisons between two independent samples and among multiple samples were performed using two-tailed t tests and one-way ANOVA, respectively. Error bars indicate at least three independent experiments, and data are shown as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.