Fig. 6

HIF1A promotes YAP1 transactivation upon nicotine treatment. a, Three potential HIF1A–binding sites (HRE1–3) in the YAP1 promoter sequence. b, The full-length and deletion mutants YAP1 promoter. c, Dual luciferase assay of YAP1 promoter activity after cotransfected the deletion mutant reporters with or without HIF1A expression vectors or control vector into Panc-1 cells. d-g, Dual luciferase assay of promoter activity in Panc-1 and BxPC3 cells after cotransfection a pGL4-HREs-luciferase plasmid and pRL-TK with HIF1A overexpression (d-e) or siRNA (f-g) vectors upon nicotine or DMSO treatment. h-i, ChIP-RT-PCR assay of the YAP promoter activity in Panc-1 (h) and BxPC3 (i) cells with or without nicotine treatment. A specific anti-HIF1A antibody was used and IgG was used as a control. The experiments were performed three times independently. HRE, hypoxia response elements; Nic, nicotine; n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001