Figure 5

P1 versus P12 intracerebral expressions of CINC-1 and levels of neutrophil infiltrations. CINC-1 was detected by ELISA at 4 and 24 h after HI in P1 brains exposed to LPS+/-HI (A). No CINC-1 was detected at 48 h after HI in P1 brains exposed or unexposed to LPS+/-HI either by ELISA (A) or IHC (data not shown) at 48 h post-HI. In contrast, following HI or LPS+HI at P12, CINC-1 was up-regulated in brains at 4, 24 and 48 h post-HI as shown by ELISA (A) and IHC experiments (B, C); this increase of CINC-1 intracerebral expression (C, arrowheads) at P12 was present in the neocortical gray matter (C), but not in the underlying white matter. Neutrophils (E, arrowheads) were detected only at P12 - not at P1 -, at 48 h post LPS+HI, in the neocortex ipsilateral to ischemia, i.e. in the areas of neocortical damage as illustrated in a schema (D). Protein detection was performed by ELISA on 3 to 4 brains assayed in duplicate at each time point under each experimental condition. IHC was performed at 48 h post-HI, in 3 to 4 brains under each experimental condition. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA with Newman-Keuls post test, except for LPS+HI versus control at P12 (48 h) by ELISA using t test with Welch correction (A).Scale bars = 15 μm (C, E).