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: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A) Immunoblot analysis of the expression and activation of selected STAT proteins in mammary epithelial cells from STAT3 and STAT5a/b conditional triple-knockout mice (MMTV-Cre Stat3/5 fl/fl ; N = 4) and age-matched Stat3/5 fl/wt controls ( N = 2 and 4, respectively). E-cadherin (CDH1) and beta-actin (ACTB) were used as loading controls. PC, wild-type control epithelial cells treated with IL-4 served as a positive control for tyrosine-phosphorylated STAT6 (pY-STAT6). (B) Immunohistochemistry of tyrosine-phosphorylated STAT1 (pY-STAT1) on histologic sections of mammary glands from postpartum STAT3/5 triple-knockout and control females; bars, 20 μm. (C and D) Immunoblot analysis of active and total levels of STAT1 before and after the retroviralbased expression of Cre recombinase in three immortalized mammary epithelial cell lines (C) and three mouse embryonic fibroblast lines (D). Wild-type mammary epithelial cells and interferon gamma-treated wild-type mouse fibroblasts served as positive controls (C) for STAT5a and active STAT1, respectively. The densitometry results of the STAT proteins in (C) from 3 technical repeats of the 3 biological repeats are shown in . (E) Immunohistochemistry of active STAT1 (pY-STAT1) in alveolar cells of mammary glands of a postpartum WAP-Cre Stat3/5 fl/fl female and an age-matched control; bars, 20 μm.

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Western Blot, Expressing, Activation Assay, Triple Knockout, Control, Positive Control, Immunohistochemistry

Journal: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A) Schematic of the knockout of STAT1, STAT3, STAT5a, and STAT5b in canonical JAK/STAT signaling cascades of the mammary gland. (B) Carmine alum-stained mammary gland wholemounts of nulliparous female mice with a targeted deletion of four Stat genes in the mammary epithelium MMTV-Cre Stat3/5 fl/fl Stat1 −/− ) and a STAT1 single-knockout littermate control; bars, 1 mm. (C) Immunoblot analysis of the seven known mammalian STAT proteins in mammary epithelial cells (MECs) from STAT1/3/5a/5b quadruple-knockout females ( N = 4) in comparison to age-matched STAT3/5a/5b triple-knockout mice ( N = 2). Other controls: C1 and C2, positive controls for active STAT3 and STAT5, mammary gland tissues from wild-type mice on day 1 of involution and lactation, respectively; C3, spleen as a positive control for STAT4; C4, wild-type MECs as a negative control for STAT4; C5 and C6 interferon-treated and untreated MECs as positive and negative controls for STAT2; C7 and C8, IL-4-treated and untreated wild-type MECs as positive and negative controls for active STAT6. GAPDH and ACTB served as loading controls.

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Knock-Out, Staining, Control, Western Blot, Quadruple Knockout, Comparison, Triple Knockout, Positive Control, Negative Control

Journal: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A) Immunoblot analysis of STAT1 and JAK1 expression and activation in response to the pharmacological inhibition of JAK1 using 1 μM itacitinib in three mammary epithelial cell lines co-deficient in STAT3, STAT5a, and STAT5b. Beta-actin (ACTB) was used as a loading control. The densitometry results of immunoblots from 2 technical repeats of the 3 biological repeats are shown in . (B) Schematic of a quadruple knockout of STAT3, STAT5a, and STAT5b along with JAK1 to genetically ablate the compensatory activation of STAT1 in the STAT3/5a/5b triple-knockout mammary epithelium and to determine the STAT-independent contribution of JAK2 to mammary gland development. (C) Carmine alum-stained mammary gland whole-mounts of 4- and 6-week-old nulliparous females that are conditionally deficient inSTAT3, STAT5a/b, and JAK1 (MMTV-Cre Stat3/5 fl/fl Jak1 fl/fl ) and littermate controls without the MMTV-Cre transgene; bars, 1 mm. Dotted lines mark the invasive fronts of the terminal ends of ducts. (D) Immunoblot analysis of STAT1 expression and activation in response to the knockout of JAK1 in the mammary epithelium of STAT3/5a/5b triple-knockout females; 4 biological repeats of quadruple-knockout females compared to 2 wild-type controls and 2 STAT3/5a/5b triple-knockout mice. Beta-actin (ACTB) served as a loading control. The densitometry results of immunoblots from the 4 biological and 2 technical repeats are shown in .

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Western Blot, Expressing, Activation Assay, Inhibition, Control, Quadruple Knockout, Triple Knockout, Staining, Knock-Out

Journal: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A and B) Immunoblot analysis of tyrosine-phosphorylated JAK1 and JAK2 and expression of selected STAT proteins in mammary epithelial cells from quadruple STAT1/3/5a/5b conditional knockout mice and wild-type controls that were treated with either oncostatin M (OSM) alone and human growth hormone (hGH) alone (A) or a combination of both (B); 2 biological repeats of experimental and control animals. GAPDH served as a loading control. (C) Immunoblot analysis to assess the activation of STAT6 and tyrosine phosphorylation of JAK1 in STAT1/3/5a/5b-deficient quadruple-knockout cells and controls following stimulation with IL-4. GAPDH was used as a loading control. PC, splenocytes as positive controls for the validated absence of STAT2 and STAT4 in the quadruple-knockout epithelial cells. (D) Summary of canonical and noncanonical signaling mechanisms by which JAK2, in cooperation with JAK1, drives the postnatal development of the mammary gland.

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Western Blot, Expressing, Knock-Out, Control, Activation Assay, Phospho-proteomics, Quadruple Knockout

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