Figure 3

Immature and mature OL loss in Timp-3 WT mice at 72 h. A. Western blots for MBP, GST-pi (π) and GalC in white matter in reperfusion injury at 72 h in WT and KO mice. No significant changes of MBP level were detected at 72 h reperfusion after ischemia in both WT and KO mice. Significantly decreased levels of GST-π was seen in ischemic WT compared with sham WT (SWT) (p < 0.01, n = 5 for each group). Significant increase of GalC was seen in ischemic WT and KO compared with sham WT (p < 0.001, n = 5 for each group) and sham KO (SKO) (p < 0.05), respectively. Timp-3 KO mouse shows significantly higher levels of GalC in white matter than WT mouse (p < 0.01). Intensity of total proteins stained by Brilliant blue R on the same blots was performed as control for protein loading and transfer normalization. The results were presented as ratio of protein expression between ischemic and nonischemic white matter. B. Double-immunostaining of GST-π (red) and GalC (green), showing the expression of mature and immature OLs in ischemic external capsule at 72 h reperfusion in Timp-3 KO and WT mice. More GST-π- and GalC-positive signals were seen in KO ischemic EC than in WT. Inserts show immunohistochemistry of GST-π and GalC in sham-operated animals. C. Co-localizarion of TUNEL labeling GST-π or GalC immunohistochemistry in ischemic WT EC at 72 h reperfusion. Arrows show the co-localization of TUNEL-positive cells with GST-π or GalC expression was seen in ischemic EC (insert 1 and 2). Scale bars = 50 μm.