Figure 7
Expression of TNF-α and TACE in ischemic EC in Timp-3 KO and WT mice at 72 h. A. Double-staining of TNF-α (red) and Iba-1 (green), showing the expression of TNF-α in microglia/macrophages (Iba-1 positive cells) in ischemic EC at 72 h reperfusion in Timp-3 KO and WT mice. More TNF-α- and Iba-1-positive cells were seen in WT ischemic EC (arrows) than in KO. Scale bar = 40 μm. B. The three-dimension (3D) and Z-stack confocal images shows the co-localization of TNF-α in macrophage. Scale bar = 5 μm. C. Quantification of Iba-1 fluorescent intensity showed greater Iba-1-positive macrophages/microglia in Timp-3 WT mice compared with KO at both 24 and 72 h (p < 0.05, n = 5 for each group). D. TACE activity measured by a fluorometric assay in ischemic white matter at 72 h reperfusion in Timp-3 KO and WT mouse brains. Significant increase in TACE relative activity was seen in ischemic WT compared with ischemic KO (p < 0.05, n = 6) and sham WT (SWT) and KO (SKO) (p < 0.01). A greater level of TACE activity was seen in nonischemic WT compared with contralateral sham WT (p < 0.05). E. Top panel: Co-localization of TACE and Iba-1 in microglia/macrophages (arrows). Bottom: TACE and CC1 co-localization in OLs (arrows). Scale bars = 50 μm. F. Z-stack confocal images show TNFR-1 co-localized in CC1-positive OLs. Western blot analysis shows no significant changes ether between ischemic and nonischemic hemispheres or between KO and WT mice (n = 6).