Figure 6
HIF-2α regulates the transcription of the MAPK3 gene, as determined by EMSA and ChIP-qPCR.
(A) Human MAPK3 promoter sequence. The “ACGTG” sequence is the putative hypoxia-response element (HRE), which may be the candidate HIF-2α binding site. The green shadow “G” site is shown relative to the transcription start site (TSS). The MAPK3 promoter probe was designed with the “ACGTG” sequence. (B) Electrophoretic mobility shift assay (EMSA) of the MAPK3 promoter probes and nuclear protein extracts. The lane to which only the biotin-labeled MAPK3 probe was added was the negative control. The anti-HIF-2α antibody was added for the supershift assay. The unlabeled MAPK3 probe was added as a competitor for the biotin-labeled probe. (C) ChIP-qPCR detection of HIF-2α binding to the MAPK3 promoter using normoxic and hypoxic hPMSCs. The tests were performed in triplicate and repeated in three independent experiments. The data are presented as the mean ± S.D. and were statistically analyzed using Student’s t-test. *p < 0.05.
