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. 2012 Jan 4;287(9):6139–6149. doi: 10.1074/jbc.M111.278630

FIGURE 6.

FIGURE 6.

FHL1 and FHL3, but not FHL2, inhibit HIF-2 transcriptional activity. A, FHL2 inhibits HIF-1, but not HIF-2 activity. 293 cells were co-transfected with p2.1, pSV-RL, HIF-1α or HIF-2α expression vector, and either EV, FHL1, FHL2, or FHL3 vector. 24 h post-transfection, the cells were lysed, and the ratio of firefly:Renilla activity was determined. B-D, FHL1–3 inhibit HIF-1 activity at target gene promoters. Hep3B cells were co-transfected with the indicated luciferase reporter, HIF-1α expression vector, and either EV, FHL1, FHL2, or FHL3 expression vector. 24 h post-transfection cells were lysed, and luciferase activity measured. E–G, FHL1 and FHL3, but not FHL2, inhibit HIF-2 activity at target gene promoters. Hep3B cells were co-transfected with the indicated luciferase reporter, HIF-2α expression vector, and either EV, FHL1, FHL2, or FHL3 expression vector. 24 h post-transfection the cells were lysed, and luciferase activity was measured. The results are shown as the means ± S.D. *, p < 0.01; #, p < 0.05 compared with EV; ns, not significant. H and I, FHL1–3 have no effect on HIF-1α or HIF-2α protein levels. 293T cells were co-transfected with HIF-1α expression vector (H) or HIF-2α expression vector (I) and either EV or V5-tagged FHL1, FHL2, or FHL3 expression vector. 24 h post-transfection the cells were lysed and probed with anti-HIF-1α or anti-HIF-2α, anti-V5, or β-actin antibodies.