Skip to main content
. 2000 Aug 1;97(16):9082–9087. doi: 10.1073/pnas.97.16.9082

Figure 5.

Figure 5

The Nip3 promoter is responsive to hypoxia and to HIF-1α. (A) The promoter sequence of Nip3 from CHO-K1 cells contains two consensus HREs (highlighted in gray). HRE1 is shown in boldface. The translation start codon of the Nip3 polypeptide at position +1 is in boldface. The 5′ end of the mRNA as determined by 5′ rapid amplification of cDNA ends is indicated by an arrow and is located 25 nucleotides downstream of a putative TATA box (underlined). HRE1-Mut changed the putative HRE beginning at position −234 from 5′-CACGTG-3′ to 5′-CACCAC-3′. HRE2-Mut changed the putative HRE beginning at position −160 from 5′-CGCGTG-3′ to 5′-CGACTG-3′. (B) Luciferase reporter constructs containing no promoter (pGL3-Basic), a minimal TATA box containing promoter (E1B-Luc), the Nip3 promoter, or the Nip3 promoter containing a mutation in the first (HRE#1-Mut) or second (HRE#2-Mut) putative HRE was transfected into 293 cells along with pcDNA3 (vector) or HIF-1α. Cells transfected with vector were incubated for 19 h in an atmosphere containing 20% O2 (normoxia) or for 5 h in an atmosphere containing 20% O2 followed by 14 h in an atmosphere containing 0.5% O2 (hypoxia). Cells transfected with HIF-1α were incubated for 19 h under normoxic (20% O2) conditions. The values represent the average luciferase activity of six samples; bars indicate standard error.