Detection of Hypoxia inducible factor-1 By DNA Binding Studies

Stabilization of HIF-1α, its translocation into the nucleus and dimerization with its HIF-1β subunit to form HIF-1 complex is an index of hypoxia. Until recently, most studies involving HIF1 were carried out in animal cell line models. In this study, we show the translocation of HIF-1 in freshly isolated human lymphocytes treated with hypoxia mimicking conditions. In order to check for expression of HIF-1 and its nuclear translocation, we used conditions of hypoxia and hypoxia mimicking conditions. in vivo hypoxia conditions were analyzed in rat models by exposing them to oxygen partial pressure relevant to an altitude of 7500 m (pO2=59 mm Hg), whereas hypoxia mimicking conditions were established by using certain agents known to stabilize HIF-1. Agents like CoCl2 and DFO were used in this context in freshly isolated lymphocytes. The translocation of HIF-1 is shown using electrophoretic mobility shift assay of nuclear and cytoplasmic extracts.