Extended Data Fig. 3: Evaluation of in vivo anti-IBD efficacy of living glues on IL-10−/− mice model.
From: Engineered living glues secrete therapeutic proteins for treatment of inflammatory bowel disease
a, Body weight changes and b, Disease activity index (DAI) recorded over 42 days post-treatments (n = 5 independent experiments, mean ± SD, a: NS (P = 0.0811), *P = 0.0422, *P = 0.0339, NS (P = 0.9161), ****P < 0.0001, b: NS (P = 0.4651), *P = 0.0249, *P = 0.0125, ****P < 0.0001). DAI scoring criteria are listed in Supplementary Table 2. c, Colon lengths (n = 5 independent experiments, mean ± s.d., ****P < 0.0001, *P = 0.0209, NS (P = 0.3138), *P = 0.0118, *P = 0.0226) and d, digital images of ex vivo gut tissues of experimental mice. e, Spleen weight (n = 5 independent experiments, mean ± s.d., ***P = 0.0002, NS (P = 0.6275), NS (P = 0.357), *P = 0.0278, *P = 0.0216) and f, digital images of ex vivo spleen tissues of experimental mice. g, Representative hematoxylin & eosin (H&E)-stained sections and h, colon damage scores of distal colon tissues from each treatment group (n = 4 independent experiments, mean ± s.d., ****P < 0.0001, NS (P = 0.0558), NS (P = 0.4651), *P = 0.0278, *P = 0.0125). The lower panel in g, shows close-up views. Scale bars = 200 and 50 μm, respectively. Scoring criteria for h are provided in Supplementary Table 3. i-j, Representative immunofluorescence staining images and k, l, m, n, semi-quantitative analysis of colon tissue sections post-treatment in mice (n = 3 independent experiments, mean ± s.d., k: ***P = 0.0002, NS (P = 0.6275), NS (P = 0.357), *P = 0.0278, ***P = 0.0003, l: ****P < 0.0001, NS (P = 0.8266), NS (P = 0.0836), **P = 0.0046, **P = 0.0023, m: ***P = 0.0001, NS (P = 0.8187), ***P = 0.0002, **P = 0.0062, **P = 0.0073, n: ***P = 0.0008, NS (P = 0.9168), NS (P = 0.0233), *P = 0.0151, ***P = 0.0046). Blue: DAPI, Red: Mucin 2, Green: E-Cadherin, Pink: ZO 1, Yellow: Occludin, Black: overlay. Scale bar = 50 μm. o, p, q, r, Levels of cytokines IL-1β, TNF, IL-6, and TGFβ from tissue homogenates, determined by ELISA (n = 5 independent experiments, mean ± s.d., o: ****P < 0.0001, NS (P = 0.6722), NS (P = 0.5671), ***P = 0.0001, ***P = 0.0002, p: ****P < 0.0001, NS (P = 0.6138), NS (P = 0.8608), *P = 0.0106, ***P = 0.0005, q: ****P < 0.0001, NS (P = 0.5395), NS (P = 0.3443), **P = 0.0045, **P = 0.0053, r: NS (P = 0.9077), NS (P = 0.8673), NS (P = 0.8097), **P = 0.0017, ***P = 0.0004). Statistical analysis was conducted using a two-tailed Student’s t-test.
