Journal: Molecular Medicine Reports

Article Title: ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis

doi: 10.3892/mmr.2025.13783

Figure Lengend Snippet: ST3Gal1 expression in inflamed IEC monolayers alters MUC2, TFF3, CDX2, STAT3 and p-STAT3 expression. ST3Gal1 knockdown in ST3Gal1-sh3/IEC significantly increased (A) MUC2, (B) TFF3 and (C) CDX2, but significantly reduced (D) STAT3 mRNA levels in the inflamed IEC monolayer. ST3Gal1 OE in ST3Gal1-OE/IEC significantly reduced (E) MUC2, (F) TFF3 and (G) CDX2 mRNA levels in the inflamed IEC monolayer. (H) ST3Gal1 OE in ST3Gal1-OE/IEC slightly decreased STAT3 mRNA levels in the inflamed IEC monolayer. (I) Western blotting showed no changes in protein expression in non-inflamed knockdown samples. (J) Western blotting showed that MUC2, TFF3 and CDX2 protein levels were increased, and p-STAT3 expression was decreased in the inflamed IEC monolayer comprising ST3Gal1-sh3/IEC. (K) Western blotting showed no notable changes in protein expression in non-inflamed OE samples. (L) Western blotting showed that MUC2, TFF3 and CDX2 protein levels were decreased, and p-STAT3 expression was increased in the inflamed IEC monolayer comprising ST3Gal1-OE/IEC. Data are presented as the mean ± SD. Statistical significance was assessed using one-way analysis of variance followed by Tukey's post-hoc test. *P<0.05 and **P<0.01. IEC, intestinal epithelial cell; sh, short hairpin; Ctr, control; OE, overexpression; MUC2, mucin 2; TFF3, trefoil factor 3; CDX2, homeobox protein CDX-2; p-, phosphorylated.

Article Snippet: Rabbit polyclonal IgG to STAT3 , Proteintech Group, Inc. , 10253-2-AP , 1:500.

Techniques: Expressing, Knockdown, Western Blot, Control, Over Expression

Journal: Molecular Medicine Reports

Article Title: ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis

doi: 10.3892/mmr.2025.13783

Figure Lengend Snippet: Comparison of the expression of inflammatory mediators (A) IL-1β, (B) IL-6 and (C) IL-8, as well as (D) STAT3 between non-lesional mucosa (n=44) and lesional mucosa (n=75) from distinct patients with ulcerative colitis. Correlations between (E) IL-1β, (F) IL-6, (G) IL-8 and (H) STAT3 mRNA levels and ST3Gal1 expression. Data were obtained from the GSE107499 dataset from the Gene Expression Omnibus database. Data in A-D are presented as box and whisker plots, and those in E-H as scatter plots. Statistical significance for group comparisons in A-D was assessed using the unpaired t-test; correlation analyses in E-H were performed via Pearson's correlation coefficient. **P<0.01.

Article Snippet: Rabbit polyclonal IgG to STAT3 , Proteintech Group, Inc. , 10253-2-AP , 1:500.

Techniques: Comparison, Expressing, Gene Expression, Whisker Assay

Journal: Poultry Science

Article Title: CSF3 enhances the innate immune responses to ALV-J infections via NF-κB and interferon pathways

doi: 10.1016/j.psj.2025.105648

Figure Lengend Snippet: CSF3 overexpression induces the expression of IFNs and antiviral genes. (A, B) Western blot analysis of env, β-actin, STAT3, and phosphorylated STAT3 (p-STAT3) protein levels in DF-1 cells infected with ALV-J following CSF3 overexpression or knockdown. (C) Quantification of env and p-stat protein levels in (A, B) based on relative grayscale values. (D, E) Western blot analysis of env, β-actin protein levels in CEF cells infected with ALV-J following CSF3 overexpression or knockdown. (F) Quantification of env protein levels in (C, D) based on relative grayscale values. (G-J) RT-qPCR analysis of mRNA expression of interferon ( IFN-α, IFN-β, IFN-γ ) and antiviral genes ( Mx, MDA5, IRF7, OASL, ACSL1, CH25H ) in DF-1 cells infected with ALV-J after CSF3 overexpression or knockdown in 12 and 24 h. (K, L) RT-qPCR analysis of mRNA expression of gp85, interferon ( IFN-α, IFN-β, IFN-γ ) and antiviral genes ( Mx, MDA5, IRF7, OASL ) in CEF cells infected with ALV-J after CSF3 overexpression or knockdown in 24 h. Statistical significance was determined using a two-tailed unpaired Student’s t-test ( p < 0.05). * p < 0.05, **p < 0.01, *** p < 0.001.

Article Snippet: Mouse Anti-ALV-J envelope protein JE9 (kindly provided by Prof. Aijian Qin, Yangzhou University, Yangzhou, China), Rabbit Anti-STAT3 antibody (bs-1141R; Boss, China; 1:1000), Rabbit Anti-phospho-STAT3 (Ser727) antibody (bs-3429R; Boss, China; 1:1000), Rabbit Anti-NFκB2 antibody (10037P, Boss, China; 1:1000), Rabbit Anti-NFκB p-p65 (bs-0982R, Boss, China; 1:1000), Rabbit Anti-IKBα Rabbit (10268-1-AP, Proteintech, USA; 1:1000), Anti-p-IKBα (bs-2513R, Boss, China; 1:1000), and goat anti-rabbit IgG/HRP (bs13278), goat anti-mouse IgG/HRP (bs12478), goat Anti-Mouse IgG ( H + L ) FITC (bs10950) secondary antibody were purchased from Bioss (Beijing, China).

Techniques: Over Expression, Expressing, Western Blot, Infection, Knockdown, Quantitative RT-PCR, Two Tailed Test

Journal: Poultry Science

Article Title: CSF3 enhances the innate immune responses to ALV-J infections via NF-κB and interferon pathways

doi: 10.1016/j.psj.2025.105648

Figure Lengend Snippet: CSF3 enhances the immune response to ALV-J via the NFκB signaling pathway. (A) Western blot analysis of p52, p100, phosphorylated p65 (p-p65), IκBα, and phosphorylated IκBα (p-IκBα) protein following CSF3 overexpression in DF-1 cells. (B) Quantification of protein levels in (A) based on relative grayscale values. (C) Western blot analysis of p52, p100, phosphorylated p65 (p-p65), IκBα, and phosphorylated IκBα (p-IκBα) protein following CSF3 knockdown in DF-1 cells. (D) Quantification of protein levels in (C) based on relative grayscale values. (D, E) RT-qPCR analysis of TNF-α, IL-1β , and IL-6 mRNA expression following CSF3 overexpression (D) or knockdown (E) in DF-1 cells. (F, G) ELISA measurement of TNF-α, IL-1β, and IL-6 protein levels following CSF3 overexpression (F) or knockdown (G) in DF-1 cells. (H, I) RT-qPCR analysis of TNF-α, IL-1β , and IL-6 mRNA expression following CSF3 overexpression (H) or knockdown (I) in CEF cells. (J, K) ELISA measurement of TNF-α, IL-1β, and IL-6 protein levels following CSF3 overexpression (J) or knockdown (K) in CEF cells. (L) Western blot analysis of STAT3, phosphorylated STAT3 (p-STAT3), env, β-actin, IκBα, p-IκBα, p-p65, p52, and p100 after STAT3 phosphorylation inhibition in CSF3-overexpressing DF-1 cells. (M) Quantification of protein levels in (L) based on relative grayscale values. Statistical significance was determined using a two-tailed unpaired Student’s t-test ( p < 0.05). * p < 0.05, **p < 0.01, *** p < 0.001.

Article Snippet: Mouse Anti-ALV-J envelope protein JE9 (kindly provided by Prof. Aijian Qin, Yangzhou University, Yangzhou, China), Rabbit Anti-STAT3 antibody (bs-1141R; Boss, China; 1:1000), Rabbit Anti-phospho-STAT3 (Ser727) antibody (bs-3429R; Boss, China; 1:1000), Rabbit Anti-NFκB2 antibody (10037P, Boss, China; 1:1000), Rabbit Anti-NFκB p-p65 (bs-0982R, Boss, China; 1:1000), Rabbit Anti-IKBα Rabbit (10268-1-AP, Proteintech, USA; 1:1000), Anti-p-IKBα (bs-2513R, Boss, China; 1:1000), and goat anti-rabbit IgG/HRP (bs13278), goat anti-mouse IgG/HRP (bs12478), goat Anti-Mouse IgG ( H + L ) FITC (bs10950) secondary antibody were purchased from Bioss (Beijing, China).

Techniques: Western Blot, Over Expression, Knockdown, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Inhibition, Two Tailed Test

Journal: Advanced Science

Article Title: Deciphering the Role and Mechanism of Decidual Monocyte‐Derived Macrophage Infiltration in Obstetric Antiphospholipid Syndrome at Single‐Cell Resolution

doi: 10.1002/advs.202503480

Figure Lengend Snippet: Abnormal infiltration of monocyte‐derived CCR2 + macrophages in OAPS decidua and their pro‐inflammatory roles. A) Bar plot showing the proportions of MDMs in decidual immune cells from OAPS patients and HCs in scRNA‐seq data. B) FCM analysis of the proportion of CCR2 + macrophages in the decidua from OAPS patients (n = 11) compared with HCs (n = 18). C) Representative images for CCR2 + and CCR2 − macrophages from the decidua of OAPS patients and HCs under the transmission electron microscope. Scale bar: 2 µm. D) Bubble plot showing the functional gene expressions in different types of decidual myeloid cells based on scRNA‐seq data. E) Bar plot showing the GO terms of marker genes in decidual MDMs. F) Bar plot showing the KEGG terms of marker genes in decidual MDMs. G) GSEA indicates the term of inflammatory response is significantly enriched in decidual CCR2 + macrophages. H) Bar plot showing the relative expression levels of genes related to inflammatory factors in decidual CCR2 + and CCR2 − macrophages, as detected using RT‐qPCR (n = 3). I) Representative immunoblots and semi‐quantified results of Arginase 1, NF‐κB P65, and STAT3 expressions in decidual CCR2 + and CCR2 − macrophages from OAPS patients (n = 3). J) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from HCs. Scale bar: 1000 µm. K) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from OAPS patients. Scale bar: 2000 µm. L) Line chart showing the effects of medium supernatants from CCR2 + and CCR2 − macrophages on HTR‐8/SVneo proliferation, as measured by CCK‐8 (n = 3). M) Bar plot showing relative expression levels of CASP3 , MMP2 , and MMP9 genes in HTR‐8/SVneo treated with supernatants from CCR2 + and CCR2 − macrophages, as detected by RT‐qPCR (n = 3). Data in (A) and (B) are presented as mean ± SD and analyzed using Student's t ‐test. Data in (H) and (I) are presented as mean ± SD and analyzed using a paired t ‐test. Data in (L) and (M) are presented as mean ± SD and analyzed using one‐way ANOVA with Dunnett's multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. OAPS, obstetric antiphospholipid syndrome; HCs, healthy controls; MDM, monocyte‐derived macrophage; FCM, flow cytometry; scRNA‐seq, single‐cell RNA sequencing; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene Set Enrichment Analysis; RT‐qPCR, real‐time quantitative polymerase chain reaction; SD, standard deviation; ANOVA, analysis of variance.

Article Snippet: The primary antibodies used: IκBα Mouse mAb (CST, USA, #4814, 1:1000), Phospho‐IκBα Rabbit mAb (CST, USA, #2859, 1:1000), Phospho‐NF‐κB p65 Rabbit mAb (CST, USA, #3033, 1:1000), NF‐κB p65 Rabbit mAb (CST, USA, #8242, 1:1000), IKKα Mouse mAb (CST, USA, #11930,1:1000), Phospho‐IKKα/β Rabbit mAb (CST, USA, #2697, 1:1000), GAPDH Mouse mAb (Proteintech, China, #60004‐1‐Ig, 1:5000), TLR4 Rabbit Polyclonal Ab (ABclonal, China, #A11226, 1:1000), Arginase‐1 Rabbit Polyclonal Ab (Proteintech, China, #16001‐1‐AP, 1:1000) and STAT3 Rabbit Polyclonal Ab (Proteintech, China, #10253‐2‐AP, 1:1000).

Techniques: Derivative Assay, Transmission Assay, Microscopy, Functional Assay, Marker, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, CCK-8 Assay, Flow Cytometry, RNA Sequencing, Real-time Polymerase Chain Reaction, Standard Deviation