Figure 7

Zebrafish phd3 induces the degradation of hif-1/2α proteins.A, Western blot analysis of hif1αa protein in HEK293T cells transfected with Flag-hif1αa together with Myc-phd3 or Myc empty vector (control) for 24 h. B and C, Western blot analysis of hif1αa protein in HEK293T cells transfected with Flag-hif1αa together with Myc-phd3 or Myc empty vector (control) for 20 h, followed by treatment with cycloheximide (CHX) (50 μg/ml) for the indicated time. The relative intensities of hif1αa in (B) were determined by normalizing the intensities of hif1αa to the intensities of β-actin. D, Western blot analysis of hif1αb protein in HEK293T cells transfected with Flag-hif1αb together with Myc-phd3 or Myc empty vector (control) for 24 h. E and F, Western blot analysis of hif1αb protein in HEK293T cells transfected with Flag-hif1αb together with Myc-phd3 or Myc empty vector (control) for 20 h, followed by treatment with CHX (50 μg/ml) for the indicated time. The relative intensities of hif1αb in (E) were determined by normalizing the intensities of hif1αb to the intensities of β-actin. G, Western blot analysis of hif2αa protein in HEK293T cells transfected with Flag-hif2αa together with Myc-phd3 or Myc empty vector (control) for 24 h. H and I, Western blot analysis of hif2αa protein in HEK293T cells transfected with Flag-hif2αa together with Myc-phd3 or Myc empty vector (control) for 20 h, followed by treatment with CHX (50 μg/ml) for the indicated time. The relative intensities of hif2αa in (H) were determined by normalizing the intensities of hif2αa to the intensities of β-actin. J, Western blot analysis of hif2αb protein in HEK293T cells transfected with Flag-hif2αb together with Myc-phd3 or Myc empty vector (control) for 24 h. K and L, Western blot analysis of hif2αb protein in HEK293T cells transfected with Flag-hif2αb together with Myc-phd3 or Myc empty vector (control) for 20 h, followed by treatment with CHX (50 μg/ml) for the indicated time. The relative intensities of hif2αb in (K) were determined by normalizing the intensities of hif2αb to the intensities of β-actin.