FIG. 3.
Cytoplasmic localization of endogenous HIF-2α in MEFs even after hypoxia treatment. Subcellular fractionation of MEFs (A) and 293 cells (C). Cells were treated or not with either 0.5% O2 hypoxia or 100 μM deferoxamine for 4 h before being harvested. (B) Immunofluorescence analysis of endogenous HIF-2α in MEFs. Wild-type MEFs were grown on the glass slide chamber and treated or not with 0.5% O2 for 4 h before fixation. HIF-2α was probed with polyclonal anti-HIF-2α antibody and detected with fluorescein isothiocyanate-labeled anti-rabbit immunoglobulin antibody.