Figure 2.

Cezanne is a positive regulator of HIF-1α protein levels

A Subcellular fractionation revealed that loss of Cezanne caused reduced HIF-1α levels in cytoplasmic (CP) and nuclear (N) fractions in both untreated and hypoxia-treated cells.

B Knockdown of Cezanne in HeLa cells subjected to hypoxia resulted in decreased HIF-1α and HIF-2α protein levels, and reduced hypoxia-induced PHD3 and BNIP3 levels compared to control cells.

C RT–PCR analysis showed no significant changes of HIF-1α mRNA levels in Cezanne-depleted cells relative to control cells.

D Overexpression of GFP-tagged wild-type (wt) or inactive (C194S) Cezanne in hypoxia-treated cells affected HIF-1α levels in a dose-dependent manner.

E Mouse embryonic fibroblasts (MEFs) isolated from Cezanne knockout mice (Cezanne^−/−) accumulated less HIF-1α in hypoxia than WT MEFs.

F Depletion of Cezanne did not affect proteins involved in the regulation of HIF-1α activity or stability.

Data information: All experiments were performed three times; bar graphs represent the mean plus standard deviation of these independent experiments.