Fig. 6: Transcription factor MXD1 binds to the promoter region of the HIF1A gene, inhibiting HIF1A transcription.

Changes in HIF-1α expression were detected by RT-qPCR (A) and western blot (B), respectively, after silencing MXD1 expression in U-251 MG and T98G cells. Values are presented as mean ± SD (n = 3), ***P < 0.001 compared to the sh-NC group. C The luciferase activity was analyzed using a dual-luciferase reporter system in 293T cells transfected with MXD1 overexpression vector and recombinant luciferase reporter vectors containing either the wild-type or MXD1 binding site mutant promoter region of the HIF1A gene. Data are presented as mean ± SD (n = 5), **P < 0.01 compared to the OE-NC group. ChIP (D) and ChIP-qPCR (E) assay was used to verify the binding site of MXD1 in the promoter region of the HIF1A gene. PCR1 represents the binding site of MXD1 in the promoter region of the HIF1A gene, while PCR2 represents the negative control group without an MXD1 binding site. Data are presented as mean ± SD (n = 3), ***P < 0.001 compared to the IgG group.