Figure 3

The fecal microbiota transplantation of IS-treated mice (FIS) induced anxiety-like behaviors and colitis in mice. FNC (fecal microbiota of control mice not treated with IS [NC]) and FIS were orally gavaged. Experimental time (a). Anxiety-like behaviors of mice were measured in the EPM (b, time spent in open arms [OT] and open arm entries [OE]), MB (marble buried, c) and LDT (d, time spent in light area [TL] and number of transitions [NT]) tasks. NF-κB (p-p65 and p65), BDNF, claudin-5, and β-actin were measured in the hippocampus by immunoblotting (e). Brain TNF-α (f) IL-1β (g) and IL-6 levels (h) were analyzed by ELISA kit. Blood corticosterone (f) IL-6 (g) and LPS levels (h) were analyzed by ELISA or LAL kit. Iba-1⁺/CD11b⁺ and CD11b⁺/CD11b⁺ populations were assayed by a confocal microscope (i). Blood corticosterone (j) TNF-α (k) IL-6 (l) and LPS levels (m) were analyzed by ELISA or LAL kit. Colitis markers colon length (n) and myeloperoxidase (MPO) activity (o) were measured in the colon. Colonic iNOS, COX-2, NF-κB (p) and tight junction proteins (occludin and claudin-1), and β-actin (q) were analyzed by immunoblotting. Fecal LPS level (r) and CD11b⁺/CD11b⁺ population (s) were analyzed by ELISA and confocal microscopy (i) respectively. FNC and FIS in figures indicate groups treated with fecal microbiota of control and IS-treated mice (suspended in 1% dextrose), respectively. All data except immunoblotting data (n = 4) were expressed as mean ± SD (n = 8). ^#p < 0.05 vs. FNC group.