Figure 3.

HIF-1:GFP protein levels are increased by hypoxia. The transgenic C. elegans strain pHJ06 Ex6, which expresses the complete HIF-1 protein fused to GFP, was harvested before (21% O₂) and after incubation in 0.1% oxygen (Upper) or 1% oxygen (Lower) for 8 h. In each experiment, a fraction of the hypoxia-treated worms were reoxygenated for 10 min (ReOx). Worms were lysed by boiling in loading buffer, and equal amounts of total protein were loaded in each lane. The expression of HIF-1:GFP and AHA-1 was assayed by probing the immunoblot with a GFP-specific antibody and the AHA-1-specific mAb 10H8.