Journal: BMC Pharmacology & Toxicology

Article Title: Network toxicology integrated with machine learning and SHAP analysis identifies overlapping immune signatures between Di(2-ethylhexyl) phthalate (DEHP) and Sjögren’s syndrome

doi: 10.1186/s40360-026-01119-x

Figure Lengend Snippet: Identification of core genes for SS classification using DEHP–SS intersect genes. ( A ) Model performance comparison heatmap showing AUC values across cohorts (generated with the R package ComplexHeatmap). ( B ) SHAP feature importance bar plot ranking genes by mean(|SHAP|). ( C ) SHAP summary (violin/beeswarm) plot showing the distribution of SHAP values per gene; point colors reflect normalized feature values. ( D ) SHAP interaction/dependence plots illustrating feature–feature interaction effects on model output. ( E ) SHAP waterfall plot explaining a representative individual prediction by decomposing feature contributions. ( F ) ROC curves for key genes (STAT1, PHGDH, ISG15, CXCL10, and CCL2). ( G ) Volcano plot of DEGs with core genes labeled

Article Snippet: Diluted STAT1 primary antibody (1:500, Bioss, bs-1317R) was added and incubated overnight at 4 °C.

Techniques: Comparison, Generated, Labeling

Journal: BMC Pharmacology & Toxicology

Article Title: Network toxicology integrated with machine learning and SHAP analysis identifies overlapping immune signatures between Di(2-ethylhexyl) phthalate (DEHP) and Sjögren’s syndrome

doi: 10.1186/s40360-026-01119-x

Figure Lengend Snippet: Molecular docking analysis of the interactions between DEHP and the core proteins. ( A ) Docking pose of DEHP with STAT1. ( B ) Docking pose of DEHP with PHGDH. ( C ) Docking pose of DEHP with ISG15. ( D ) Docking pose of DEHP with CXCL10. ( E ) Docking pose of DEHP with CCL2. Docking indicates binding propensity under the scoring function and does not constitute experimental validation

Article Snippet: Diluted STAT1 primary antibody (1:500, Bioss, bs-1317R) was added and incubated overnight at 4 °C.

Techniques: Binding Assay, Biomarker Discovery

Journal: BMC Pharmacology & Toxicology

Article Title: Network toxicology integrated with machine learning and SHAP analysis identifies overlapping immune signatures between Di(2-ethylhexyl) phthalate (DEHP) and Sjögren’s syndrome

doi: 10.1186/s40360-026-01119-x

Figure Lengend Snippet: Single-cell atlas localization of STAT1 in submandibular gland tissue. ( A ) t-SNE plot of the PanglaoDB submandibular gland single-cell dataset (SRA693675:SRS3206192; Mus musculus). ( B ) Feature plot showing STAT1 expression enrichment in salivary mucous cells

Article Snippet: Diluted STAT1 primary antibody (1:500, Bioss, bs-1317R) was added and incubated overnight at 4 °C.

Techniques: Single Cell, Expressing

Journal: BMC Pharmacology & Toxicology

Article Title: Network toxicology integrated with machine learning and SHAP analysis identifies overlapping immune signatures between Di(2-ethylhexyl) phthalate (DEHP) and Sjögren’s syndrome

doi: 10.1186/s40360-026-01119-x

Figure Lengend Snippet: Regulatory effect of DEHP on STAT1 expression in HSG cells. ( A ) Western blot showing representative STAT1 and GAPDH bands from the same membrane/exposure. ( B ) Quantification of relative STAT1 protein expression (* p < 0.05, ** p < 0.01). ( C ) RT-qPCR analysis of relative STAT1 mRNA expression (* p < 0.05, ** p < 0.01). ( D ) Immunofluorescence staining of STAT1 (green) and nuclei (DAPI, blue) in HSG cells treated with different concentrations of DEHP

Article Snippet: Diluted STAT1 primary antibody (1:500, Bioss, bs-1317R) was added and incubated overnight at 4 °C.

Techniques: Expressing, Western Blot, Membrane, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Gut Microbes

Article Title: Simulated microgravity induces cerebral dysfunction by disturbing protective microbiota-metabolite-microglia signaling across the gut‒brain ax is

doi: 10.1080/19490976.2026.2635820

Figure Lengend Snippet: LA inhibits the activation of inflammatory microglia by binding to STAT1 and blocking its phosphorylation at Tyr 701 and Ser 727 sites. A. Schematic of the experimental design in vitro . BV2 microglial cells were cultured under SMG for 72 h, followed by the treatment with LA. After 24 h post-LA administration, cells were collected for RNA sequencing and other detection assays. B. Cell viability was determined by CCK8 assay with the exposure of LA at indicated concentrations for 24 h: 0 μM vs 150 μM, p = 0.0286; 0 μM vs 200 μM, p = 0.0286. C. RNA sequencing heatmap analysis of BV2 disease-associated microglia makers in SMG-LA (−), SMG-LA (50) (50 μM), or SMG-LA (100) (100 μM) group. D. RNA sequencing heatmap analysis of BV2 M1/M2 makers in SMG-LA (−), SMG-LA (50) (50 μM), or SMG-LA (100) (100 μM) group. E. (i) Immunofluorescent analysis for IBA1 and CD68 expression in BV2 (scale bars, 100 μm); (ii) Statistics on the mean fluorescence intensity of E(i). NG vs SMG-LA (−), p < 0.0001; SMG-LA (−) vs SMG-LA (50) (50 μM), p < 0.0001. F. Detection of reactive oxygen species (ROS) levels using DCFH-DA probe in BV2 cells. (i) Immunofluorescent analysis; (ii) Statistics on the mean fluorescence intensity of F(i). NG vs SMG-LA (−), p = 0.0002; SMG-LA (−) vs SMG-LA (50) (50 μM), p < 0.0001. Rosup as a positive control. G. (i) Immunoblot analysis of three independent individuals for t-STAT1, p -STAT1(Tyr 701) and p -STAT1(Ser 727) in the hippocampus. (ii) Relative protein expression of STAT1 (NG vs SMG-LA (−), p = 0.0071; SMG-LA (−) vs SMG-LA (50) (50 μM, p = 0.0031) and p -STAT1(Tyr 701) (NG vs SMG-LA (−), p = 0.0111; SMG-LA (−) vs SMG-LA (50) (50 μM, p = 0.0004) by grayscale analysis. H. (i) Immunoblot analysis of three independent samples for STAT1, p -STAT1(Tyr 701) and p -STAT1(Ser 727) in BV2 cells. (ii) Relative protein expression of STAT1 (NG vs SMG-LA (−), p = 0.0448; SMG-LA (−) vs SMG-LA (50) (50 μM, p = 0.001), p -STAT1(Tyr 701) (NG vs SMG-LA (−), p = 0.0448; SMG-LA (−) vs SMG-LA (50) (50 μM, p = 0.0168) and p -STAT1(Ser 727) (NG vs SMG-LA (−), p = 0.0065; SMG-LA (−) vs SMG-LA (50) (50 μM, p = 0.0007) by grayscale analysis. I. Molecular docking diagrams of LA binding to STAT1. J. Surface plasmon resonance (SPR) analysis of the interaction between LA and STAT1. Results are based on three independent biological replicates. Data are shown as the mean ± SD (rats’ sample) or mean ± SEM (BV2 sample). The analysis is performed using two-tailed unpaired Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns no significance.

Article Snippet: STAT1 Polyclonal antibody , Proteintech , 10144-2-AP.

Techniques: Activation Assay, Binding Assay, Blocking Assay, Phospho-proteomics, In Vitro, Cell Culture, RNA Sequencing, CCK-8 Assay, Expressing, Fluorescence, Positive Control, Western Blot, SPR Assay, Two Tailed Test

Journal: Animal Bioscience

Article Title: Transcriptomic analysis identifies CXCL12 as a novel candidate gene for litter size in rabbits

doi: 10.5713/ab.24.0640

Figure Lengend Snippet: CXCL12 and its receptor CXCR4 were expressed and localized in the ovary, with their binding activating the JAK/STAT signaling pathway. (A) Immunofluorescence staining of ovarian follicles revealed that CXCL12 was co-localized with its receptor CXCR4 in ovarian follicles. (B) CXCL12 overexpression significantly promoted the expression of CXCR4 , JAK2 and STAT1 . (C) Knocking down CXCL12 significantly inhibited the expression of CXCR4 , JAK2 and STAT1 . (D) CXCL12 overexpression in GCs promoted the protein expression of CXCR4, JAK2 and STAT1 as well as the phosphorylation of JAK2 and STAT1. When CXCL12 was knocked down, the results were reversed. (E) MSX-122 inhibitor-based treatment further confirmed that CXCL12 promoted the total protein and phosphorylation levels of JAK2 and STAT1 (The t test was used for the above analyses comparing two individual samples. * p<0.05; ** p<0.01; *** p<0.001).

Article Snippet: Protein detection was achieved using the following antibodies: anti-CCND1 mouse monoclonal antibody (1:250, Proteintech), anti-PCNA rabbit polyclonal antibody (1:250, Proteintech), anti-Bcl2 rabbit polyclonal antibody (1:250, Proteintech), anti-Bax rabbit polyclonal antibody (1:250, Proteintech), anti-CITED1 rabbit polyclonal antibody (1:50, Proteintech), anti-WNT10B mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-CXCR4 mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-phospho-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam, Cambridge, UK), anti-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam), anti-phospho-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-GAPDH mouse monoclonal antibody (1:2,500, Proteintech), 1:1,000 goat anti-rabbit secondary antibody IgG (Proteintech) and 1:1,000 goat anti-mouse secondary antibody IgG (Proteintech).

Techniques: Binding Assay, Immunofluorescence, Staining, Over Expression, Expressing, Phospho-proteomics

Journal: Animal Bioscience

Article Title: Transcriptomic analysis identifies CXCL12 as a novel candidate gene for litter size in rabbits

doi: 10.5713/ab.24.0640

Figure Lengend Snippet: CXCL12 and its receptor CXCR4 were expressed and localized in the ovary, with their binding activating the JAK/STAT signaling pathway. (A) Immunofluorescence staining of ovarian follicles revealed that CXCL12 was co-localized with its receptor CXCR4 in ovarian follicles. (B) CXCL12 overexpression significantly promoted the expression of CXCR4 , JAK2 and STAT1 . (C) Knocking down CXCL12 significantly inhibited the expression of CXCR4 , JAK2 and STAT1 . (D) CXCL12 overexpression in GCs promoted the protein expression of CXCR4, JAK2 and STAT1 as well as the phosphorylation of JAK2 and STAT1. When CXCL12 was knocked down, the results were reversed. (E) MSX-122 inhibitor-based treatment further confirmed that CXCL12 promoted the total protein and phosphorylation levels of JAK2 and STAT1 (The t test was used for the above analyses comparing two individual samples. * p<0.05; ** p<0.01; *** p<0.001).

Article Snippet: Protein detection was achieved using the following antibodies: anti-CCND1 mouse monoclonal antibody (1:250, Proteintech), anti-PCNA rabbit polyclonal antibody (1:250, Proteintech), anti-Bcl2 rabbit polyclonal antibody (1:250, Proteintech), anti-Bax rabbit polyclonal antibody (1:250, Proteintech), anti-CITED1 rabbit polyclonal antibody (1:50, Proteintech), anti-WNT10B mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-CXCR4 mouse monoclonal polyclonal antibody (1:250, Proteintech), anti-phospho-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam, Cambridge, UK), anti-JAK2 rabbit monoclonal polyclonal antibody (1:250, Abcam), anti-phospho-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-STAT1 rabbit polyclonal antibody (1:250, Proteintech), anti-GAPDH mouse monoclonal antibody (1:2,500, Proteintech), 1:1,000 goat anti-rabbit secondary antibody IgG (Proteintech) and 1:1,000 goat anti-mouse secondary antibody IgG (Proteintech).

Techniques: Binding Assay, Immunofluorescence, Staining, Over Expression, Expressing, Phospho-proteomics

Journal: Blood Advances

Article Title: STAT1-mediated epigenetic regulation of LIN28A controls iPSC-derived platelet production through the let-7–RALB axis ∗

doi: 10.1182/bloodadvances.2024015557

Figure Lengend Snippet: STAT1 is the upstream regulator of LIN28A in imMKCLs. The siRNA SMARTpool designed against human STAT1 (siSTAT1) and a siNT were transfected into imMKCLs. (A) A schematic illustration of the experimental procedure of siRNA-mediated STAT1 knockdown in imMKCLs. (B) STAT1 knockdown significantly decreased STAT1 and LIN28A mRNA expression and increased let-7a-5p expression. (C) Intracellular protein levels of STAT1 (mean fluorescence intensity detected in siNT or siSTAT1 imMKCLs by intracellular flow cytometry. STAT1 expression was compared using a histogram overlay in the left panel. (D) STAT1 knockdown improved iPSC-PLT generation by imMKCLs (clone 7). (E) Representative flow cytometry plots of iPSC-PLTs generated from siNT or siSTAT1 imMKCLs in static or turbulent flow conditions. CD41 + CD42b + platelets were identified from a platelet-sized gate. (F) STAT1 knockdown decreased mRNA expression of representative immune-related genes in imMKCLs. (G) The secretion of IL-8 from clone 7 and clone 7-3 during the maturation stage (DOX-OFF day 6). Data are expressed as the mean ± SEM from 3 to 4 independent experiments. Unpaired 2-tailed Student t tests were used to assess statistical significance. IL-8, interleukin 8; MK, megakaryocyte; ns, not significant.

Article Snippet: After 2 washes with phosphate-buffered saline, cells were incubated with an anti-STAT1 polyclonal antibody (Proteintech; 10144-2-AP; 0.4 μg/100 μL) for 1 hour on ice.

Techniques: Transfection, Knockdown, Expressing, Fluorescence, Flow Cytometry, Generated

Journal: Blood Advances

Article Title: STAT1-mediated epigenetic regulation of LIN28A controls iPSC-derived platelet production through the let-7–RALB axis ∗

doi: 10.1182/bloodadvances.2024015557

Figure Lengend Snippet: Pharmacological inhibition of STAT1 enhances iPSC-PLT production under turbulent flow conditions. (A) Chemical structures of fludarabine and flavopiridol, the 2 established inhibitors for STAT1 phosphorylation. (B) Dose optimization on iPSC-PLT production of fludarabine (0 to ∼5 μM) and flavopiridol (0 to ∼100 nM). (C) iPSC-PLT generation by imMKCLs (clone 7) in the absence and presence of fludarabine (1 μM) or flavopiridol (10 nM) under static or turbulent flow conditions. (D) Representative flow cytometry plots of iPSC-PLTs generated in the absence and presence of fludarabine (1 μM) or flavopiridol (10 nM) in static or turbulent flow conditions. CD41 + CD42b + platelets were identified from a platelet-sized gate. (E) Fludarabine (1 μM) or flavopiridol (10 nM) treatments elevated let-7a-5p expression. (F) Flavopiridol (10 nM) treatment reduced mRNA expression of representative immune-related genes in imMKCLs at the maturation stage. Data are expressed as the mean ± SEM from 4 to 5 independent experiments. Statistical significance was assed using 1-way analysis of variance or unpaired 2-tailed Student t tests. DMSO, dimethyl sulfoxide.

Article Snippet: After 2 washes with phosphate-buffered saline, cells were incubated with an anti-STAT1 polyclonal antibody (Proteintech; 10144-2-AP; 0.4 μg/100 μL) for 1 hour on ice.

Techniques: Inhibition, Phospho-proteomics, Flow Cytometry, Generated, Expressing