Skip to main content
. 2017 Jun 30;292(34):13970–13985. doi: 10.1074/jbc.M117.778134

Figure 8.

Figure 8.

Effect of hypoxia on CHGA promoter haplotype 2: Crucial role for c-Rel. A, role of c-Rel in haplotype-specific activation of CHGA promoter Hap2 in response to hypoxia. CHGA promoter Hap2–GLuc and Hap5–GLuc constructs were transfected into IMR-32 cells with/without c-Rel expression plasmid and subjected to 12 h of hypoxia. Results are expressed as mean ± S.E. of triplicate values of Gaussia luciferase/μg of protein, *, p < 0.05; **, p < 0.01; and ****, p < 0.0001 when compared with basal activity of the construct and ###, p < 0.001 and ####, p < 0.0001 when compared with the corresponding treatment in the CHGA promoter Hap5 construct. B, effect of hypoxia on c-Rel expression. IMR-32 cells were subjected to hypoxia for 12 h and Western blotting of the total proteins was carried out probing for HIF-1α, c-Rel, and GAPDH. C and D, effect of hypoxia on the binding of c-Rel to −1018T and −57T alleles. Chromatin immunoprecipitation of N2a cells transfected with CHGA promoter Hap2 or Hap5 constructs, with/without exposure to hypoxia was carried out using antibody against c-Rel/preimmune IgG. Immunoprecipitated chromatin was subjected to qPCR and fold-enrichment in the signal obtained from the c-Rel antibody over preimmune IgG is shown. *, p < 0.05 when compared with fold-enrichment in untreated conditions and #, p < 0.05; ##, p < 0.01; ###, p < 0.001; and ####, p < 0.0001 as compared with the corresponding treatment in case of CHGA promoter Hap2 construct.