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. 2021 Jun 7;9:675486. doi: 10.3389/fcell.2021.675486

FIGURE 3.

FIGURE 3

ATF4 bound the Ihh gene promoter and promoted its expression of human NP cells. (A) Two predicted sites (S1 and S2) that ATF4 bound on the promoter region of Ihh. (B) AGE indicated two putative DNA sequences of Ihh immunoprecipitated by the anti-ATF4-coded beads. (C) Dual-luciferase reporter assay in NP cells co-transfected with the reporter plasmid and ATF4. Structural schematic of the wt and mutant sequences of S1 and S2 in the human Ihh promoter region is shown. Luciferase activity driven by the wt2 Ihh promoter was more pronounced following ATF4 treatment, and it was not driven by the mut2 luciferase reporter upon ATF4 treatment. In contrast, luciferase activity driven by the wt2 luciferase reporter decreased in the absence of ATF4. Luciferase activity showed no difference when driven by wt1 luciferase reporter. Data are presented as means ± SD (n = 3). ***p < 0.001, - indicates not applied, and + indicates applied. ATF4, activating transcription factor 4; Ihh, Indian hedgehog; AGE, agarose gel electrophoresis; wt, wild type.