Journal: Journal of Inflammation Research

Article Title: Downregulation of IFIT3 Relieves the Inflammatory Response in Ulcerative Colitis via Selectively Regulating Macrophage M1 Polarization and the STAT1/2 Signaling Pathway

doi: 10.2147/JIR.S542033

Figure Lengend Snippet: IFIT3 knockdown relieved the intestinal inflammatory response and inhibited the activation of the STAT1/2 pathway in DSS-induced UC mice. ( A and B ) Comparison of colon transduction efficacy in mice between intraperitoneal injection of saline and AAV9 using paraffin sections ( A ) and the IVIS Lumina III imaging system ( B ). Scale bar = 100 µm. ( C ) Comparison of colon length between AAV9-shNC- and AAV9-shIFIT3-treated UC mice. ( D ) Weight changes in the two mouse groups. ( E ) Representative images and histological score of H&E staining in colon tissues from the two mouse groups. Scale bar = 100 µm (top) and 25 µm (bottom). ( F ) mRNA levels of IFIT3, TNF-α, IL-1β and IL-6 in colon tissues from the two mouse groups. ( G ) Protein expressions of STAT1, pSTAT1, STAT2, pSTAT2 and IFIT3 in colon tissues from the two mouse groups. Data was presented as mean ± SD (n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The PVDF membranes were incubated with primary antibodies against IFIT3 (1:3000; #15201-1-AP, Proteintect), STAT1 (1:1000; # R25799 , Zenbio, Chengdu, China), pSTAT1 (1:1000; #bsm-52209R, Bioss, Beijing, China), STAT2 (1:1000; #R381419 Zenbio), pSTAT2 (1:1000; #bs-3428R, Bioss), and GAPDH (1:1000; #GB15002-100, Servicebio) overnight at 4 °C and then incubated with HRP-labeled secondary antibodies at room temperature for 1 hour.

Techniques: Knockdown, Activation Assay, Comparison, Transduction, Injection, Saline, Imaging, Staining

Journal: Journal of Inflammation Research

Article Title: Downregulation of IFIT3 Relieves the Inflammatory Response in Ulcerative Colitis via Selectively Regulating Macrophage M1 Polarization and the STAT1/2 Signaling Pathway

doi: 10.2147/JIR.S542033

Figure Lengend Snippet: IFIT3 knockdown repressed macrophage M1 polarization by inhibiting the STAT1 pathway in vitro. ( A and B ) mRNA and protein levels of IFIT3 in untreated THP‐1 macrophages or treated with 100 ng/mL LPS or 100 ng/mL LPS + 20 ng/mL IFN‐γ or 20 ng/mL IL‐4. ( C ) mRNA levels of IFIT3 in 100 ng/mL LPS-stimulated macrophages treated with siNC or two siRNAs targeting IFIT3 (siIFIT3-1 and siIFIT3-2). ( D ) mRNA levels of TNF-α, IL-1β and IL-6 in 100 ng/mL LPS-stimulated macrophages treated with siNC and siIFIT3-2. ( E ) Protein levels of STAT1, pSTAT1, STAT2, pSTAT2 and IFIT3 in each group. ( F ) Representative gating and the co-expression of CD86 and CD68 in untreated (M0) and 100 ng/mL LPS-stimulated macrophages (M1) by flow cytometry. ( G ) Representative gating and the co-expression of CD68 and CD86 in 100 ng/mL LPS-stimulated macrophages treated with siNC and siIFIT3-2 by flow cytometry. Data was presented as mean ± SD from 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The PVDF membranes were incubated with primary antibodies against IFIT3 (1:3000; #15201-1-AP, Proteintect), STAT1 (1:1000; # R25799 , Zenbio, Chengdu, China), pSTAT1 (1:1000; #bsm-52209R, Bioss, Beijing, China), STAT2 (1:1000; #R381419 Zenbio), pSTAT2 (1:1000; #bs-3428R, Bioss), and GAPDH (1:1000; #GB15002-100, Servicebio) overnight at 4 °C and then incubated with HRP-labeled secondary antibodies at room temperature for 1 hour.

Techniques: Knockdown, In Vitro, Expressing, Flow Cytometry

Journal: Journal of Inflammation Research

Article Title: Downregulation of IFIT3 Relieves the Inflammatory Response in Ulcerative Colitis via Selectively Regulating Macrophage M1 Polarization and the STAT1/2 Signaling Pathway

doi: 10.2147/JIR.S542033

Figure Lengend Snippet: UPA inhibits inflammation by silencing IFIT3 in LPS-stimulated macrophages. ( A ) mRNA levels of IFIT3, TNF-α, IL-1β and IL-6 in 100 ng/mL LPS-stimulated macrophages with untreated or treated with 100 nM and 400 nM UPA. ( B ) Protein levels of STAT1, pSTAT1, STAT2, pSTAT2 and IFIT3 among cell-groups. ( C ) Schematic diagram of relation among UPA, IFIT3 and STAT1/2 pathways. Data was presented as mean ± SD from 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The PVDF membranes were incubated with primary antibodies against IFIT3 (1:3000; #15201-1-AP, Proteintect), STAT1 (1:1000; # R25799 , Zenbio, Chengdu, China), pSTAT1 (1:1000; #bsm-52209R, Bioss, Beijing, China), STAT2 (1:1000; #R381419 Zenbio), pSTAT2 (1:1000; #bs-3428R, Bioss), and GAPDH (1:1000; #GB15002-100, Servicebio) overnight at 4 °C and then incubated with HRP-labeled secondary antibodies at room temperature for 1 hour.

Techniques:

Journal: bioRxiv

Article Title: Gut microbial metabolites butyrate and acetate limit Zika virus replication and associated ocular manifestations via the G-protein coupled receptor 43/FFAR2

doi: 10.1101/2025.07.15.664962

Figure Lengend Snippet: HTMCs (n=3) were pretreted with PBA, NaB, or NaAc followed by ZIKV (Z) (PRVABC59 strain, MOI: 1) infection for 48h. Mock-treated cells were used as controls. The cell lysates from mock, ZIKV-infected, and drug-treated cells were subjected to western blotting for the NFκB, ERK1/2, p38, STAT1, STAT2, STST3, RIG-I, IRF3, and IFIT2 pathways. (B) Densitometric analysis of the immunoblots was performed using ImageJ. The bar graph represents means ± SD from three biological replicates. * P < 0.05; ** P < 0.005; *** P < 0.0005; ****, P < 0.0001 (one-way ANOVA).

Article Snippet: Antibodies used in this study were purchased from the following sources: 4G2 (GeneTex, #GTX57154), NS3 (GeneTex, #GTX133309), β-actin (Millipore Sigma, #A2228), pNFκB (#3033), NFκB (#6956) pERK1/2 (#4370), ERK (#4695), pP38 (#4511), P38 (#8690), pSTAT1 (#9167), STAT1 (#14994), pSTAT2 (#88410), STAT2 (#72604), pSTAT3 (#9145), STAT3 (#9139), RIG-I (#3743), pIRF3 (#79945), IRF3 (#4302), and IFIT2 (#92633) antibodies were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Infection, Western Blot

Journal: bioRxiv

Article Title: Gut microbial metabolites butyrate and acetate limit Zika virus replication and associated ocular manifestations via the G-protein coupled receptor 43/FFAR2

doi: 10.1101/2025.07.15.664962

Figure Lengend Snippet: IFNAR1 −/− mice (n=6-8) were pretreated with FFAR2 inhibitor 4-CMTB followed by PBA, NaB, or NaAc via i.p. administration, and ZIKV infection as per the timeline shown in . Seven days post-infection, anterior segment/TM tissue from treated and untreated mice were harvested and subjected to (A) RNA extraction and qPCR to measure the mRNA expression levels of PRRs (RIG-I, TLR3), inflammatory (IL-6, IL-1β, CCL-4), IFNs (IFN-α2, IFN-β1), and ISGs (ISG15, OAS2, MX1), and (B) western blot for NFκB, MAPKs (ERK1/2, p38), STAT1, STAT3, RIG-I, and IRF7 inflammatory/ISG pathways. (C) Densitometric analysis of the immunoblots was performed using ImageJ. The bar graph represents means ± SD from three biological replicates. * P < 0.05; ** P < 0.005; *** P < 0.0005; ****, P < 0.0001 (one-way ANOVA).

Article Snippet: Antibodies used in this study were purchased from the following sources: 4G2 (GeneTex, #GTX57154), NS3 (GeneTex, #GTX133309), β-actin (Millipore Sigma, #A2228), pNFκB (#3033), NFκB (#6956) pERK1/2 (#4370), ERK (#4695), pP38 (#4511), P38 (#8690), pSTAT1 (#9167), STAT1 (#14994), pSTAT2 (#88410), STAT2 (#72604), pSTAT3 (#9145), STAT3 (#9139), RIG-I (#3743), pIRF3 (#79945), IRF3 (#4302), and IFIT2 (#92633) antibodies were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Infection, RNA Extraction, Expressing, Western Blot