Fig. 6. Association of BNIP3 with subcellular fractions. Cardiac myocytes were subjected to hypoxia as described in Fig. 1. At the indicated times, cells were harvested, rinsed and lysed (Kubasiak et al., 2002). Lysed cells were untreated (left panels) or subjected to alkaline solubilization (right) (Kubasiak et al., 2002). The pH of each sample was verified by sampling small aliquots. After treatments, samples were separated into subcellular fractions and analyzed by western blots. Blots were stripped and re-probed with anti-succinate dehydrogenase (Upstate Biotechnology, Lake Placid, NY, USA) probes to define the purity of fractions. Results are representative of two separate experiments.