Figure 4

Caspase-3 expression in ischemic EC in Timp-3 KO and WT mice at 24 and 72 h. A. Immunostaining shows that expression of caspase-3 was detected in ischemic WT EC 24 h after reperfusion. The increased caspase-3 expression in WT was present in OL-like cells. Less caspase-3 immunostaining was detected in both ischemic WT and KO ECs at 72 h, while nuclear counterstaining with DAPI (blue) showed greater loss of cells in ischemic WT EC. Insert 1 and 2 show caspase-3 immunostaing is associated with nuclei (arrows). Scale bars = 50 μm. B. Western blots for expression of caspase-3 in white matter at 24 h in Timp-3 KO and WT mice. The graph shows an increase in pro-caspase-3 band at 32 kDa in the ischemic EC of the WT mice (p < 0.05, n = 4). Cleaved caspase-3 (17 kDa and 19 kDa) was not detected. Brilliant blue R staining on the same blots was performed as control for protein loading and transfer. C. Western blots for caspase-3 in white matter in reperfusion injury at 72 h showed no difference in expression of pro-caspase-3 between WT and KO, but increased cleaved caspase-3 (19 kDa and 17 kDa) in ischemic WT white matter (p < 0.05, n = 6 for each group).