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. 2004 Apr 8;23(9):1949–1956. doi: 10.1038/sj.emboj.7600196

Figure 2.

Figure 2

HIF-1α induction of p21cip1 and G1 arrest is independent of its DNA-binding and transcriptional activity. (A) Transcriptional activity of HIF-1α, ΔODD, and ΔODD plus C800V or LCLL mutations was determined in Cos-7 cells in a HIF-1-mediated reporter system (Huang et al, 1996). Relative luciferase units (RLU) are presented with means plus standard errors from three independent experiments. (B, D) HCT116 cells were assayed for VEGF expression by RT–PCR (B) or VEGF secretion by ELISA (D) after adenoviral infection as indicated. DFO, desferrioxamine. (C, E) ΔODD LCLL mutant was analyzed for its effects on p21cip1 expression at mRNA (C) and protein (E) levels. (F) Effect of the ΔODD LCLL mutant on cell cycle profile was determined as above, with the percentage of each phase indicated in the inlet. (G–I) ΔODD and its R27G mutant were assayed in Cos-7 cells for HIF-1-mediated reporter activity (G) and Western analysis of protein levels (H), and in HCT116 cells for induction of cell cycle arrest (I). CON, transfected with pcDNA3.