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. 2016 Apr 25;47(5):1354–1363. doi: 10.1161/STROKEAHA.115.011800

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© 2016 The Authors.

Stroke is published on behalf of the American Heart Association, Inc., by Wolters Kluwer. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDervis License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 1. — Experimental setup and infarct volume. A, Experimental setup (3 independent experiments). Experimental groups: microbiota-depleted AB(+/−) MCAo/sham, with antibiotic treatment stopped 72 h before surgery; microbiota-depleted AB(+/+) MCAo/sham, with antibiotic treatment during the entire experiment; recolonized AB(+/−) MCAo/sham, microbiota-depleted mice recolonized with SPF microbiota; antibiotic treatment was stopped 48 h before recolonization; microbiota-depleted AB(+/−) naïve, microbiota-depleted animals without any surgical intervention with antibiotic cocktail stopped 72 h before the experiment; SPF AB(−/−) MCAo, conventionally colonized (specific pathogen-free microbiota) mice without any antibiotic treatment, subjected to MCAo surgery; SPF AB(+/−) MCAo, SPF mice with antibiotic treatment for 48 h up to 72 h before surgery. B, Infarct volume assessed by MRI at day one after MCAo did not differ between investigated groups. Microbiota-depleted AB(+/−) n=25, microbiota-depleted AB(+/+) n=9, recolonized AB(+/−) (microbiota-depleted recolonized with SPF microbiota) n=7, SPF AB(+/−) (SPF with antibiotic treatment for 48 h) n=8, SPF AB(−/−) n=18. Box plot with whiskers minimum to maximum. No statistically significant differences were found when comparing all experimental groups (Kruskal Wallis test with Dunn’s post hoc) or when comparing microbiota-depleted AB(+/−) with SPF AB(−/−) mice Mann–Whitney test). FACS indicates flow cytometric analysis; MCAo, middle cerebral artery occlusion; MLN, mesenteric lymph nodes; MRI, magnetic resonance imaging; PP, Peyer’s patches; and SPF, specific pathogen-free.