Fig. 6.
Increased caspase 3 and calpain activation in Mobr mice after mild neonatal hypoxic/ischemic insult. (A) Mice undergoing a mild hypoxic/ischemic insult were killed 24 hr after injury, and the activity of caspase 3 was analyzed by examining the cleavage of 7-amino-4-methylcoumarin, N-acetyl-l-aspartyl-l-glutamyl-l-valyl-l-aspartic acid amide (Ac-DEVD-AMC) (see Materials and Methods) in the left injured (L) and right uninjured (R) hippocampal lysates from wild-type (+/Y, n = 14) and Mobr (−/Y, n = 11) littermates. (B) Lysates from injured (L) and uninjured (R) hippocampus of wild-type and Mobr (−/Y) littermates were subjected to SDS/PAGE, transferred to Immobilon P membranes, and stained with primary antibody against spectrin and the caspase 3-dependent p120 cleavage product, as well as the p145 and p150 calpain-dependent cleavage products. The lower band reveals this same membrane reprobed with antibody against tubulin as loading control. KO, knockout.