Figure 6.

Hypoxia potentiates HIF-1α-regulated CASC9 transcriptionally. (A) SW1990 and PANC-1 cells were incubated under the hypoxic (1%) condition for different durations, and the mRNA level of CASC9 was measured by qRT-PCR. (B) Two cell lines were treated with 100 μM CoCl₂ for 24 h, and qRT-PCR was used to detect the expression of CASC9. Two cell lines were transfected with Si-HIF-1α or Si-NC, followed by incubation under the 1% hypoxic condition for 9 h, and (C) the protein expression of p-AKT and HIF-1α was determined by western blot analysis; (D) the mRNA level of CASC9 was measured by qRT-PCR. (E) The putative binding sites of HIF-1α to the promoter of CASC9 are shown. (F) After treatment with 1% hypoxia for 9 h, a ChIP assay was carried out to determine the changes in binding abilities of different putative binding sites. The graphs shown are representative results from three independently repeated experiments. The mRNA value was normalized to GAPDH. **, P < 0.01; ***, P < 0.001.