Loss of cell attachment to the extracellular matrix causes apoptosis, a process known as anoikis. BCL2 is an antiapoptotic protein that can protect cells against anoikis. BIT1 is evolutionarily conserved from bacteria to human. It contains an N-terminal mitochondria localization sequence and a C-terminal UPF0099 domain. Northern blot analysis detected a 1-kb BIT1 transcript expressed prominently in testis, prostate, skeletal muscle, and liver, with lower expression in heart, spleen, placenta, and colon. No significant expression was detected in thymus or peripheral leukocytes. Human embryonic kidney cells showed only trace expression of BIT1. Immunostaining localized endogenous BIT1 to the mitochondria of HeLa cells, and immunoblot analysis revealed a 20-kD doublet in HeLa cell mitochondrial extracts.