Figure 5.

Reoxygenation reverses acute hypoxia-induced PKG1 ubiquitin modification. FPASMCs were exposed to hypoxia for 2 hours and transferred to normoxia for an additional 2 hours. Cell lysates were immunoprecipitated with PKG1 antibody and analyzed according to Western blotting, using anti-ubiquitin monoclonal antibody, and were reprobed with PKG1 antibody. The experiment was repeated three times, and a representative blot is shown. Duration of treatment time in hours is indicated. H, hypoxia (3% O₂); N, normoxia (21% O₂). Hypoxic cells that were reoxygenated contained no ubiquitinated PKG1 (Ub-PKG), similar to normoxic cells that had never been exposed to hypoxia.