Journal: Open Life Sciences

Article Title: AUDA suppresses apoptosis of HCAECs induced by TNF and serum of Kawasaki disease patients by inhibiting endoplasmic reticulum stress

doi: 10.1515/biol-2025-1254

Figure Lengend Snippet: AUDA inhibits ERS by suppressing the JAK/STAT1 pathway. (A) 45 mmol/L of STAT1 transcription enhancer (2-NP) was used to activate the JAK/STAT1 signaling pathway. (B) Cell viability was detected using CCK8 assay. *** P < 0.001 versus TNF control, &&& P < 0.001 versus TNF and AUDA-treated cells. (B) The level of LDH was detected using Elisa assay. (C) The expression of ERS marker factors was detected. The protein levels of IRE1α (D), XBP1 (E), CHOP (F), ATF6 (G), Cleaved-Caspase12 (H) and Caspase12 (I) were standardized to GAPDH. (J) Heatmap of ERS marker factors expression. *** P < 0.001.

Article Snippet: 45 mmol/L of STAT1 transcription enhancer (2-NP) (MedChemExpress, Monmouth Junction, NJ, USA) was used to activate the Janus kinase (JAK)/STAT1 signaling pathway.

Techniques: CCK-8 Assay, Control, Enzyme-linked Immunosorbent Assay, Expressing, Marker

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were blocked with 5 % non-fat milk in TBS-T and incubated overnight at 4 °C with primary antibodies against phospho-AKT (Thr308), phospho-NFκB p65 (Ser536), phospho-p38 MAPK (Thr180/Tyr182), phospho-STAT1 (Tyr701), and phospho-STAT3 (Tyr705) (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were stripped and re-probed for total AKT, NFκB p65, p38 MAPK, STAT1, STAT3, and β-actin (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA) as a loading control.

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Redox Biology

Article Title: Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis

doi: 10.1016/j.redox.2026.104096

Figure Lengend Snippet: HDCA-treated ameliorate inflammation and modulate lipid metabolism in vivo. (A) Acetyl-CoA levels decreased over time in HDCA-treated Tregs. (B – E) Quantitative real-time PCR analysis of the mRNA expression of key lipogenic genes, including (B) fatty acid synthase (FASN), (C) acetyl-CoA carboxylase (ACC), (D) SCD1, and (E) SREBP-1c, in Tregs with or without HDCA treatment. (F) Functional metabolic flux assay using 13 C-glucose tracing shows decreased enrichment of labeled β-hydroxypalmitate in the HDCA-treated group. (G) Immunoblot analysis of phosphorylated STAT1 in Tregs following HDCA exposure, and quantification statistical analysis results are presented in the bar graphs. (H) Representative IHC images show reduced IL-21 expression in atherosclerotic lesions of mice following HDCA treatment (magnification, 5 × ; scale bar, 50 μm). (I) Plasma cholesterol levels were measured by enzymatic colorimetry assay in FXR-sufficient controls and FXR KO mice ± HDCA. (J) Plasma triglyceride levels in the Vector and FXR KO groups were quantified at 0, 6, 12, and 18 h by a colorimetric assay. (K) Hemodynamic profiling assessed heart rate, systolic blood pressure (SBP), and diastolic blood pressure (DBP) for each group. (L – M) Ratios of monounsaturated to saturated fatty acids (MUFA/SFA) and polyunsaturated to saturated fatty acids (PUFA/SFA) in serum cholesterol esters (CE) and triglycerides (TG) were quantified by lipidomics in FXR-sufficient control and FXR KO mice ± HDCA. Data are presented as the mean ± SD (n = 3-5 biological replicates). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Proteins were detected using the following antibodies: anti-CPT1a antibody (ab234111, abcam), anti-beta actin antibody (ab8226, abcam), anti-PERK antibody (ab229912, abcam), anti-ERK1+ERK2 antibody (ab184699, abcam), anti-S6K1 antibody (ab14708, abcam), anti-S6K1 (phospho T229) antibody (ab5231, abcam), Rac1/2/3 antibody (G-2) (sc-514583, Santa Cruz), anti-Calpain 1 antibody (ab108400, abcam), anti-MMP2 antibody (ab92536, abcam), anti-IL-10 antibody (ab310329, abcam), anti-ZNF671 antibody (HPA046099, Sigma-Aldrich), anti-MAPK6/ERK3 antibody (ab53277, abcam), SIAH1 recombinant rabbit monoclonal antibody (PSH01-80) (MA5-51926, Thermo Fisher), p-Stat1 antibody (pY701.4A) (sc-136229, Santa Cruz), Stat1 antibody (C-136) (sc-464, Santa Cruz), anti-FXR1 antibody (ab155124, abcam), phospho-Raptor (Ser792) polyclonal antibody (PA5-118730, Thermo Fisher), anti-PD1 antibody (ab214421, abcam), SHP-2 antibody (3752S, Cell Signaling Technology), IL-10R antibody (3F9) (sc-53654, Santa Cruz), GAPDH antibody (6C5) (sc-32233, Santa Cruz), rabbit anti-mouse IgG H&L (HRP) (ab6728, abcam), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody, Alexa FluorTM Plus 488 (A32731, Thermo Fisher).

Techniques: In Vivo, Real-time Polymerase Chain Reaction, Expressing, Functional Assay, Flux Assay, Labeling, Western Blot, Clinical Proteomics, Colorimetric Assay, Plasmid Preparation, Control

Journal: bioRxiv

Article Title: STAT1 expression in myeloid cells restrains murine norovirus-induced hepatitis and fibrosis

doi: 10.64898/2026.04.26.720966

Figure Lengend Snippet: Stat1 Het and Stat1 KO littermates were i.v. infected with 2x10 5 PFU of CR6 (or sham-infected) and analyzed at 7 days pi (A-F) or indicated timepoints (G-I). (A, B) Percent weight change, n=4 naive and n=15-16 MNV CR6-infected Stat1 Het and Stat1 KO . (C) Whole liver and spleen at d7 pi. (D) Representative images of H&E-stained, FFPE liver sections. (E) Hepatic MNV CR6 genome copies detected by qRT-PCR, normalized to tissue weight. (F) Quantified areas of necroinflammation in liver sections. (D-F) n=4 naive and n=10 MNV CR6-infected Stat1 Het and Stat1 KO . (G, H) Representative images (G) and quantified areas of necroinflammation (H) in H&E-stained, FFPE liver sections of naive and CR6-infected Stat1 KO (n=5-8 per timepoint). (I) RNAscope detection of MNV CR6 genomes and DNA (DAPI). All graphs are pooled from at least two independent experiments with n=4 naive and n=15-16 MNV CR6-infected Stat1 Het and Stat1 KO . Images are representative of two independent experiments. Mann-Whitney U test with Holm-Šídák correction for multiple comparisons where applicable. ** = p < 0.01, **** = p < 0.0001, ns = not significant. Scale bars = 100 µm unless otherwise indicated. LoD = limit of detection.

Article Snippet: MNV-free C57BL/6J (000664), Stat1 KO (B6.129S(Cg)- Stat1 tm1Dlv /J, 012606), and BoyJ (B6.SJL- Ptprc a Pepc b , 002014) mice were sourced from Jackson Laboratories and bred in-house.

Techniques: Infection, Staining, Quantitative RT-PCR, RNAscope, MANN-WHITNEY

Journal: bioRxiv

Article Title: STAT1 expression in myeloid cells restrains murine norovirus-induced hepatitis and fibrosis

doi: 10.64898/2026.04.26.720966

Figure Lengend Snippet: Stat1 Het and Stat1 KO mice were i.v. infected with 2x10 5 PFU of CR6 (or sham-infected) and analyzed at 5 days pi (A-D) or the times indicated (E-H). All flow-based measurements represent cells recovered from the left lateral lobe of the liver. (A) Quantification of CR6 genome copies in flow-sorted CD45 - and CD45 + cells. n=4-5 per group from 1 independent trial. (B) Quantification of live CD45 + leukocytes isolated from the liver left lateral lobe. (C) Total numbers of hepatic CD45 + immune populations, represented in the heatmap as the log 2- fold change relative to uninfected controls (CD19 + TCRβ - B cells, CD19 - TCRβ + CD8α + T cells, myeloid lineage cells all pre-gated on CD19 - TCRβ -: CD11b + F4/80 - CD11c - Ly6C + Ly6G - monocytes; CD11b + F4/80 + MΦ, CD11b + F4/80 - CD11c - Ly6C + Ly6G + neutrophils, F4/80 - CD11c + DCs). (D) RT-qPCR quantification of CR6 genome copies in flow-sorted cell populations (gating strategy as in C). Data in B-D are compiled from 2 independent experiments with n=4 naive, n=7 Stat1 Het , and n=10 Stat1 KO CR6-infected mice. (E) Naive or CR6-infected Stat1 KO FFPE liver sections stained for CD11b, MNV NS1, and DNA (DAPI) (top) or Masson’s Trichrome (bottom). Scale bars = 100 µm. Representative images from 2 independent experiments. (F, G) Quantification of live CD45 + leukocytes, and CD45 + CD19 - TCRβ - CD11b + F4/80 + MΦ recovered from the left lateral lobe of naive or CR6-infected Stat1 KO livers. (H) Gene expression of ccl2, tnfa, and ifng in liver RNA, normalized to hprt expression, expressed as fold-change compared to uninfected controls. Data are pooled from two independent experiments, n=5-7 per group. Mann-Whitney U test, with Holm-Šídák correction for multiple comparisons where applicable (B, F, H). * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001, ns = not significant. Scale bars = 100 µm. LoD = limit of detection.

Article Snippet: MNV-free C57BL/6J (000664), Stat1 KO (B6.129S(Cg)- Stat1 tm1Dlv /J, 012606), and BoyJ (B6.SJL- Ptprc a Pepc b , 002014) mice were sourced from Jackson Laboratories and bred in-house.

Techniques: Infection, Isolation, Quantitative RT-PCR, Staining, Gene Expression, Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: STAT1 expression in myeloid cells restrains murine norovirus-induced hepatitis and fibrosis

doi: 10.64898/2026.04.26.720966

Figure Lengend Snippet: Stat1 Het and Stat1 KO mice were i.v. infected with 2x10 5 PFU of CR6 (n = 5 mice per group). At 5 days pi, RNA was extracted from the left lateral lobe of the liver for sequencing (A-F). (A) Volcano plot of all analyzed genes. Data points are coloured if p adj < 0.05 and log 2 fold change > or < 2. (B) GSEA of published liver disease gene sets (autoimmune hepatitis: GSE9892, IL-1β injection: GSE216278, alcohol toxicity: GSE214778, and fibrosis: GSE222576) significantly enriched in Stat1 KO mice. (C) Genes from a published analysis of human acute Hepatitis B liver failure (GSE62037) were converted to mouse IDs (see methods). The heatmap represents genes enriched in both acute Hepatitis B liver failure and CR6-infected Stat1 KO mice (NES = 2.99, padj < 0.00001). (D) GSEA was performed against Gene Ontology Biological Process terms. Selected significant terms differentially up- or down-regulated in Stat1 KO mice are shown. (E) Heatmap of the Macrophage Activation GO term (NES = 1.90, p adj = 0.0002) arranged by unsupervised hierarchical clustering. (F) GSEA comparison between CR6-infected Stat1 KO liver tissue and published gene sets from BMDM activated by LPS & IFNγ, IL-4 & IL-13, or both (GSE138263). (G-H) Stat1 Het and Stat1 KO BMDM were infected with 5x10 5 PFU of CR6, and RNA was extracted 24 hours pi. (G) RT-qPCR quantification of CR6 genome copies in cultured BMDM. (H) Gene expression of il1b and arg1 in CR6-infected BMDM, normalized to hprt expression, and expressed as fold-change relative to uninfected controls. (I) Relative expression of AAMΦ signature genes arg1 and chil3 in flow-sorted CD45 + cells isolated from the left lateral lobe of CR6-infected Stat1 Het and Stat1 KO mice at 5 days pi, normalized to hprt expression and uninfected controls. Data are pooled from two (H), three (G), or one (I) independent experiments. Mann-Whitney U-test. ** = p < 0.01, *** = p < 0.001. NES = normalized enrichment score. LoD = limit of detection.

Article Snippet: MNV-free C57BL/6J (000664), Stat1 KO (B6.129S(Cg)- Stat1 tm1Dlv /J, 012606), and BoyJ (B6.SJL- Ptprc a Pepc b , 002014) mice were sourced from Jackson Laboratories and bred in-house.

Techniques: Infection, Sequencing, IF-P, Injection, Activation Assay, Comparison, Quantitative RT-PCR, Cell Culture, Gene Expression, Expressing, Isolation, MANN-WHITNEY

Journal: bioRxiv

Article Title: STAT1 expression in myeloid cells restrains murine norovirus-induced hepatitis and fibrosis

doi: 10.64898/2026.04.26.720966

Figure Lengend Snippet: STAT1 expression in hematopoietic cells is necessary and sufficient for protection from CR6-induced liver disease . (A) Experimental design for bone marrow chimera experiments. Lethally irradiated Stat1 KO (CD45.2 + ) and WT (BoyJ, CD45.1 + ) mice received i.v. bone marrow transplants from sex-matched Stat1 KO or WT donors. After 6 weeks of reconstitution, mice were infected with 2x10 5 PFU of CR6 and analyzed at day 7 pi. (B) Percent expression of donor congenic CD45 marker on PBMCs before infection. (C) Percent weight change from initial over time. † indicates euthanasia due to humane endpoint. (D) TCID 50 quantification of infectious CR6 in liver lysate normalized to tissue weight. (E) Quantification of hepatic CR6 genome copies, normalized to tissue weight. (F, G) H&E staining (F) and (G) quantified areas of necroinflammation in FFPE liver sections of CR6-infected mice. (H) Masson’s trichrome staining of FFPE livers at 7 days post CR6-infection. Scale bars = 100 µm. Data are pooled from two independent experiments (B-C, n=7-17 per group), or are representative of two independent experiments (D-G). For (C), the extra sum-of-squares F test was performed on fifth-order polynomial lines fitted to both groups or to each group individually. For all other significance testing, Mann-Whitney U-test with Holm-Šídák correction for multiple comparisons. * = p < 0.05, **** = p < 0.0001, ns = not significant. LoD = limit of detection.

Article Snippet: MNV-free C57BL/6J (000664), Stat1 KO (B6.129S(Cg)- Stat1 tm1Dlv /J, 012606), and BoyJ (B6.SJL- Ptprc a Pepc b , 002014) mice were sourced from Jackson Laboratories and bred in-house.

Techniques: Expressing, Irradiation, Infection, Marker, Staining, MANN-WHITNEY

Journal: bioRxiv

Article Title: STAT1 expression in myeloid cells restrains murine norovirus-induced hepatitis and fibrosis

doi: 10.64898/2026.04.26.720966

Figure Lengend Snippet: Bone marrow chimeras were generated as in and analyzed at day 16 post-CR6 infection. Stat1 KO → Stat1 KO chimeras reached humane endpoint at day 7 pi, precluding analysis at the later timepoint. (A) Quantification of hepatic CR6 genome copies, normalized to tissue weight. (B) Total numbers of CD45 + immune populations isolated from the left lateral lobe of the liver, represented in the heatmap as the log 2- fold change relative to uninfected controls (CD19 + TCRβ - B cells, CD19 - TCRβ + CD8α + T cells, myeloid lineage cells all pre-gated on CD19 - TCRβ - non-B/non-T cells: CD11b + F4/80 - CD11c - Ly6C + Ly6G - monocytes; CD11b + F4/80 + MΦ, CD11b + F4/80 - CD11c - Ly6C + Ly6G + neutrophils, F4/80 - CD11c + dendritic cells). (C, D) H&E or Masson’s trichrome staining of FFPE livers at day 16 pi. (E) Expression of fibrosis-related genes chil3 and mmp9 in the liver of CR6-infected chimeras, normalized to hprt expression and expressed as fold-change relative to uninfected controls. Scale bars = 100 µm. Data are representative of two independent experiments with n=2-7 mice per group. LoD = limit of detection.

Article Snippet: MNV-free C57BL/6J (000664), Stat1 KO (B6.129S(Cg)- Stat1 tm1Dlv /J, 012606), and BoyJ (B6.SJL- Ptprc a Pepc b , 002014) mice were sourced from Jackson Laboratories and bred in-house.

Techniques: Generated, Infection, Isolation, Staining, Expressing