Figure 2.

Sp1 and HIFs contribute to the synergistic activation of multiple genes in OVSAYO cells under SSH. a) Synergistic activation of genes under SSH. N and H indicate normoxia (20% O₂ for 16 h) and hypoxia (1% O₂ for 16 h), respectively. Data shown represent the mean (n = 3) ± SD. b) Activation of multiple genes under hypoxia is dependent on HIFs. Western blotting is also shown for HIFs. Data shown are the mean (n = 3) ± SD. c) The ICAM1 promoter region and predicted Sp1 binding sites (ref. 19). PCR-amplicons generated in the reporter gene assay are shown in Additional file 2: Figure S1a. Bold bars: mutated sites (Sp1-mut 1 and 2); underlined nucleotide sequences: regions deleted by mutagenesis. d, e) Luciferase reporter gene assay. Plasmids were co-transfected with the indicated vectors, and luciferase activities were measured at 24 h post-transfection. Luciferase activities measured in cells transfected with the empty vector were defined as 100%. Data shown are the mean (n = 3) ± SD. *P = 0.0018; **P = 0.075.