Figure 8

Figure 8

Zebrafish phd3 promotes polyubiquitination of hif-1/2α proteins.A, the ubiquitination of hif1αa protein in HEK293T cells transfected with the indicated plasmids for 24 h, followed by MG-132 (20 μM) treatment for 8 h. The relative intensities of hif1αa ubiquitination were determined by normalizing the intensities of poly-ubiquitinated hif1αa in the Ni²⁺-pulldown to the intensities of hif1αa in the total cell lysates (TCL). B, the ubiquitination of hif1αb protein in HEK293T cells transfected with the indicated plasmids for 24 h, followed by MG-132 (20 μM) treatment for 8 h. The relative intensities of hif1αb ubiquitination were determined by normalizing the intensities of poly-ubiquitinated hif1αb in the Ni²⁺-pulldown to the intensities of hif1αb in the TCL. C, the ubiquitination of hif2αa protein in HEK293T cells transfected with the indicated plasmids for 24 h, followed by MG-132 (20 μM) treatment for 8 h. The relative intensities of hif2αa ubiquitination were determined by normalizing the intensities of poly-ubiquitinated hif2αa in the Ni²⁺-pulldown to the intensities of hif2αa in the TCL. D, the ubiquitination of hif2αb protein in HEK293T cells transfected with the indicated plasmids for 24 h, followed by MG-132 (20 μM) treatment for 8 h. The relative intensities of hif2αb ubiquitination were determined by normalizing the intensities of poly-ubiquitinated hif2αb in the Ni²⁺-pulldown to the intensities of hif2αb in the TCL.