Mcl-1 is abundantly expressed in the testis, and Mcl-1 levels do not change following ischemia or ischemia-reperfusion. (A) Results of Mcl-1 immunoblot analysis of nuclear proteins isolated from experimentally-induced unilaterally ischemic testes (I) and sham-treated testes (S). Aliquots containing 75-μg of nuclear proteins were analyzed from testes subjected to 1 h or 6 h of ischemia, and ischemia and reperfusion (I/R) for 1h/4h. Mcl-1 was detected at ~40kDa. Extracts from the PC-12, Ramos and BJAB cell lines were used as positive controls for detecting Mcl-1. Activating transcription factor-2 (ATF-2) was detected as a loading control as a protein that is constitutively expressed and is unaffected by ischemia. (B) Histogram showing Mcl-1 quantitation normalized to ATF-2 as a percentage of the sham controls. No statistically significant differences in the steady state levels of the Mcl-1 protein were observed between the ischemic and normoxic groups (ANOVA, p<0.05, n=3-5).