Journal: Investigative Ophthalmology & Visual Science

Article Title: Gut Microbial Metabolite Indole-3-Acetic Acid Exerts Protective Effects by Regulating Orbital Fibroblast Fibrosis in Thyroid Eye Disease

doi: 10.1167/iovs.67.10.35

Figure Lengend Snippet: Activation of AHR alleviated fibrotic phenotypes by regulating THE TGF-β1/Smad2/3 signaling pathway. ( A , B ) Protein expression of fibronectin, collagen Iα1, α-SMA, AHR, and CYP1A1, and the phosphorylation of Smad2 and Smad3 in TED OFs treated with IAA and CH-223191 detected using a western blotting assay ( n = 3 in each group). ( C ) Immunofluorescence assay measured the distribution of AHR in TED OFs ( n = 3 in each group). Blue represents DAPI, and red represents AHR. Scale bar : 20 µm. ( D , E ) Protein expression of fibronectin, collagen Iα1, α-SMA, AHR, and CYP1A1, and the phosphorylation of Smad2 and Smad3 in non-TED OFs treated with IAA and CH-223191 detected using a western blotting assay ( n = 3 in each group). ( F ) Immunofluorescence assay measured the distribution of AHR in non-TED OFs. ( n = 3 in each group). Blue represents DAPI, and red represents AHR. Scale bar : 20 µm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 compared to the TGF-β1 group (one-way ANOVA).

Article Snippet: To evaluate the role of AHR signaling in OF activation, cells were pretreated with 10-μM AHR antagonist CH-223191 (MedChemExpress) for 2 hours before stimulation with TGF-β1 and IAA.

Techniques: Activation Assay, Expressing, Phospho-proteomics, Western Blot, Immunofluorescence

Journal: Investigative Ophthalmology & Visual Science

Article Title: Gut Microbial Metabolite Indole-3-Acetic Acid Exerts Protective Effects by Regulating Orbital Fibroblast Fibrosis in Thyroid Eye Disease

doi: 10.1167/iovs.67.10.35

Figure Lengend Snippet: IAA attenuated cell migration by activating AHR in OFs. ( A – D ) TED OFs ( A , B ) and non-TED OFs ( C , D ) were treated with TGF-β1, IAA, and the AHR antagonist CH-223191 for 24 or 48 hours. Cell migration was examined using a scratch wound healing assay ( n = 3 in each group). Scale bar : 100 µm. * P < 0.05, *** P < 0.001, **** P < 0.0001 compared to the TGF-β1 group (one-way ANOVA).

Article Snippet: To evaluate the role of AHR signaling in OF activation, cells were pretreated with 10-μM AHR antagonist CH-223191 (MedChemExpress) for 2 hours before stimulation with TGF-β1 and IAA.

Techniques: Migration, Wound Healing Assay

Journal: bioRxiv

Article Title: Helminth-remodeled microbial indole-3-lactic acid drives AhR-dependent disease tolerance

doi: 10.64898/2026.05.06.723221

Figure Lengend Snippet: (A) Mice were orally infected with 500 TsML, followed by oral administration of vehicle control or L. murinus (1 × 10^9 CFU/mL) from 1 to 14 dpi, and at 14 dpi, adult worms were recovered from each group to assess the burden of T. spiralis (n=6). (B-D) In vivo effects in antibiotic-treated mice. Western blot analysis and quantification of AhR and Cyp1a1 protein levels in the duodenum after supplementation with vehicle, L. murinus (1×10 9 CFU/mL ) for 7days (n=3). (E-L) Flow cytometric analysis of Treg (CD4 + CD25 + Foxp3 + ) and Th17 (CD4 + IL-17A + ) cells in mesenteric lymph nodes (E, F, G and H) and spleen (I, J, K and L) after supplementation with vehicle, L. murinus (1×10 9 CFU/mL) for 7days after 21 days of antibiotic pretreatment (n=5). (M) Mice were orally infected with 500 TsML, followed by oral administration of vehicle control or ILA (20 mg/kg) from 1 to 14 dpi, and at 14 dpi, adult worms were recovered from each group to assess the burden of T. spiralis (n=6). (N-P) In vivo effects in antibiotic-treated mice. (N-P) Western blot analysis and quantification of AhR and Cyp1a1 protein levels in the duodenum after supplementation with vehicle, ILA (20 mg/kg) for 7days (n=3). (Q-X) Flow cytometric analysis of Treg (CD4 + CD25 + Foxp3 + ) and Th17 (CD4 + IL-17A + ) cells in mesenteric lymph nodes (Q, R, S and T) and spleen (U, V, W and X) after supplementation with vehicle, ILA (20 mg/kg) for 7days after 21 days of antibiotic pretreatment (n=5). (Y) Representative immunofluorescence images confirming increased AhR and Cyp1a1 protein in organoids treated with L. murinus (1×10 6 CFU/mL) or ILA (2 mM) for 48 h. Data are representative of three independent experiments. Data are presented as mean ± SD and were analyzed by Student’s t-test (A, C, D, F, H, J, L, M, O, P, R, T, V, and X). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: For the AhR antagonist pretreatment experiments, mice were intraperitoneally injected with 10 mg/kg of CH223191 (Med Chem Express) for 14 days while being infected with T. spiralis .

Techniques: Infection, Control, In Vivo, Western Blot, Immunofluorescence