Skip to main content
. 2014 May 19;9(5):e98136. doi: 10.1371/journal.pone.0098136

Figure 5. The HRE is required for anoxia-induced MUC5AC transcription.

Figure 5

(A) Chromatin immunoprecipitation (ChIP) assay. NCI-H292 cells were transfected with vectors encoding HIF-1α-binding flanking region of the MUC5AC promoter (HRE site), or a region that does not contain an HRE region (non-HRE site; negative control). The cells were treated under normoxic or anoxic conditions for 6 h and subjected to immunoprecipitation with HIF-1α antibody or control IgG. (B) The MUC5AC mRNA transcript was analyzed by real-time PCR. The binding to HRE under anoxic conditions significantly increased the expression of MUC5AC expression compared to non-HRE region, or HRE region under anoxic conditions (n = 3, *p<0.05). (C) EMSA to determine HIF-1α binding to the HRE region of the MUC5AC promoter in response to anoxia. The activity with HIF-1α-specific HRE-containing oligonucleotides is increased remarkably in response to anoxia. The EMSA band of interest was found to be selectively inhibited by specific HRE-containing competitor oligonucleotide, and it was super-shifted by anti-HIF-1α antibody. Data is shown as mean ± standard deviation. (N; normoxia, A; anoxia).