Fig. 3.

Effects of minocycline on microglia/macrophage phagocytosis post SAH. Mice were operated on day 0 and treated with 45 mg/kg/dose with minocycline or PBS over 14 days. Then acute brain slices were prepared, treated, and stained, and images were recorded, as indicated in the Methods section. a Microglia/macrophage phagocytosis was significantly increased because of SAH and reflected by the accumulation of fluorescent beads (green) in Iba1-positive cells [46]. Minocycline significantly reduced phagocytic activity of microglia/macrophages, as depicted by a lower amount of fluorescent beads co-localized with Iba1-positive cells. Representative images were analyzed using ImageJ software. Relative number of fluorescent beads co-localized with Iba1-positive cells (b), the relative amount of phagocytic active microglia/macrophages (c), and the number of phagocytic active microglia/macrophages assigned to the analyzed brain area relatively to the ACA puncture (d) are depicted in the graphs. Bar indicates 20 µm. Values from all graphs are means ± SEM (n = 5 animals per group). ****^/####P < 0.0001, ***^/###P < 0.001, **^/##P < 0.01, and *^/#P < 0.05 versus sham + vehicle and SAH + vehicle, respectively. Statistical significance was determined by one-way ANOVA Bonferroni corrected. (*) describes significance with respect to sham; (^#) describes significance with respect to the SAH + vehicle. ACA anterior cerebral artery, ANOVA analysis of variance, DAPI 4',6-diamidino-2-phenylindol, Iba1 ionized calcium-binding adapter molecule 1, PBS phosphate buffered saline, SAH subarachnoid hemorrhage, SEM standard error of mean (Color figure online)