Figure 5

ICH-induced gelatinase activation is compromised in TLR2 KO mice. (A-B) At 24 h post-ICH, mice were sacrificed and cryosections were incubated in fluorescein-conjugated DQ gelatin for 2 h. Fluorescence due to gelatinase activity in the perihematoma region was visualized under fluorescent microscope. Scale bars: 50 μm. (C-D) Following in situ zymography, sections from WT mice were immunostained with anti-GFAP (C) or NeuN (D) antibodies. The locations of the gelatinase activity shown in the panels C and D are denoted in panel E. Scale bars: 25 μm. (F) At 6 h post-collagenase injection, total RNA was isolated from ipsilateral hemorrhagic tissue of WT and TLR2 KO mice (n = 4) and used to measure MMP2 and MMP9 mRNA levels. Data are presented as mean ± SEM (** p < 0.01, vs. collagenase-injected WT mice). (G-J) Brain sections of WT or TLR2 KO ICH mice (n = 3) were immunostained with anti-MMP9 antibody 24 h after collagenase injection. (K-P) ICH injured brain sections obtained from WT or TLR2 KO mice were co-stained with anti-MMP9 and anti-GFAP (K-M) or anti-CD68 (N-P) antibodies to determine the cellular source of MMP9. Scale bars: 50 μm.