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. 2023 Aug 9;16(6):1011–1031. doi: 10.1016/j.jcmgh.2023.08.002

Figure 2.

Figure 2

Supplementation of D-aa ameliorate experimental colitis. (A–D) We co-housed C57BL/6J mice from the same parents until 7 weeks old before the experiment started. We then divided the mice into cages with 2 mice in each and randomly administered D-aa/L-aa. Seven-week-old C57BL/6J mice received drinking water with 1% 5-mix L-aa (L-aa5mix-WTDSS) or D-aa (D-aa5mix-WTDSS) or sterile water (Cont-WTDSS) for 3 weeks prior to 7 days of 2% DSS challenge. BW change (A), DAI score (B), H&E-stained histological sections of the colon (C), and histological scores (D) are shown. N = 14, 8, and 16 (A, B); N = 8, 8, and 8 (C, D) (Cont-WTDSS, L-aa5mix-WTDSS, D-aa5mix-WTDSS, respectively). (E–I) We co-housed Rag2-/- mice from the same parents until 7 weeks old before the experiment started. We then divided the mice into cages with 2 mice and randomly administered D-aa. Rag2-/- mice aged 7 weeks received drinking water with or without 1% D-aa5mix (Cont-Rag2-/-DSS, D-aa5mix-Rag2-/-DSS) for 3 weeks prior to 7 days 2% DSS challenge. BW change (E), percentage of BW on day 8 (F), the DAI score (G), H&E-stained histological sections from the colon (H), and histological score (I) are shown. N = 8, 14 (E, F); N = 7, 8 (G); N = 8, 8 (H, I) (Cont-Rag2-/-DSS and D-aa5mix- Rag2-/-DSS, respectively). (J–N) We co-housed Myd88-/- mice from the same parents until 7 weeks old before the experiment started. We then divided the mice into cages with 2 mice in each and randomly administered D-aa. Myd88-/- mice aged 7 weeks received drinking water 1% D-aa 5MIX (D-aa5mix-Myd88-/-DSS, n = 8), sterile water (Cont-Myd88-/-DSS, n = 7) for 3 weeks prior to 7 days 2% DSS challenge. BW change (J), percentage of BW on day 6 (K), the DAI score (L), H&E-stained histological sections of the colon (M), and histological score (N) are shown. N = 7, 8 (J–L); N = 4, 8 (M, N) (Cont-Myd88-/-DSS and D-aa5mix-Myd88-/-DSS, respectively). (O–R) We co-housed Rag2-/- mice from the same parents until 7 weeks old before the experiment started. We then divided the mice into cages with 2 mice in each and randomly administered D-aa. Rag2-/- mice aged 7 weeks transferred naive CD4+CD45RBhigh T cells received drinking water with or without 1% D-aa 5-mix for 4 weeks (T-D-aa5mix-Rag2-/-, n = 3; T-Cont-Rag2-/-, n = 3). BW change (O), fecal lipocalin-2 (LCN-2) (P), H&E-stained histological sections from colon (Q), and histological score (R) are shown. (S–U) Seven-week-old GF C57BL/6J mice received a fecal transplant from WT mice treated with or without 5-mix D-aa (ContFMT, n = 8; D-aaFMT, n = 9) following 2% DSS in drinking water. BW change (S), percentage of BW on day 7 (T), and the DAI score (U) are shown. (V, W) PCoA plots of unweighted UniFrac distance (V) and LDA scores (W) of the fecal microbiome obtained from Cont-WTDSS and D-aa5mix-WTDSS mice in Figure 2 (A). All experiments were performed in at least 2 replicates (n = 1–6, combined). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. P values were obtained via the Student t-test and analysis of variance with Tukey’s post hoc test (A, B, D). Data are shown as standard error of the mean.