Cellular localization of three cell adhesion molecules (E-selectin, VCAM-1, and ICAM-1) expression and co-expression of E-selectin, VCAM-1, and ICAM-1 with KLF4 on cerebral blood vessels after focal cerebral ischemia. Images show the dual-IF staining for E-selectin/VCAM-1/ICAM-1 with CD31, Mac-1, GFAP, or KLF4 in ischemic hemisphere at day 2 post-ischemia. Scale bar = 100 μm (inserts = 40 μm). Note that E-selectin/VCAM-1/ICAM-1 always co-localized with CD31-positive vessels, especially on the leaked blood vessels. In addition, E-selectin/VCAM-1/ICAM-1 co-localized with quite a few Mac-1-positive microglia/inflammatory macrophages in the ischemic hemisphere. Furthermore, E-selectin/VCAM-1/ICAM-1 also co-localized with GFAP-positive astrocytes in the penumbra (penumbra = P), but in the ischemic core (core = C), this was markedly reduced. Of interest, a fraction of E-selectin/VCAM-1/ICAM-1 co-localized with KLF4, but the distribution patterns of their expressions were different, where the high levels of KLF4 were expressed, relatively low levels of E-selectin/VCAM-1/ICAM-1 were expressed on the blood vessels in the ischemic hemisphere, and vice versa.