Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: bioRxiv

Article Title: Neural precursor cells rescue symptoms of Rett syndrome by activation of the Interferon γ pathway

doi: 10.1101/2024.01.07.574507

Figure Lengend Snippet: A) Principal component analysis (PCA) plot for the sequenced samples of WT, Mecp2 KO, WT+NPCs and Mecp2 KO+NPCs cerebella. Percentage of variance is reported for both PC1 (first component) and PC2 (second component). B) The number of deregulated genes (DEGs) for the different comparison is reported, considering a p.adj<0.05. The number of DEGs with a LogFoldChange lower than −1 (down-regulated genes) or LogFoldChange greater than 1 (up-regulated genes) is also indicated. C) Dot plot of Gene Ontology (GO) enriched pathway analysis in the cerebellum, indicating the top 30 most enriched pathways of the comparison between KO+NPCs versus KO samples. D) Gene set enrichment analysis (GSEA) of IFNγ response in cerebellar samples, indicating a significant enrichment of the gene set in KO+NPCs vs KO comparison, as well as in KO+NPCs vs WT comparison. E) The histogram reports the transcriptional levels of genes (Parp9, Iftm3, Irf8 and Bst2) associated to IFNγ pathway. Data, expressed as percentage of WT animals, are shown as mean ± SEM. *p<0.05, **p<0.01 by two-way ANOVA followed by Tukey post hoc test. F) Western blot analysis of phosphorylated STAT1 (P-STAT1) in WT or KO neurons cultured with NPCs (for 14 days) (n=7-9). Data are represented as mean ± SEM and expressed as percentage of WT neurons cultured alone. Representative bands of P-STAT1 and the corresponding lanes of TGX-stain free gel are reported.

Article Snippet: Then, membranes were incubated for 1 hour in blocking solution [5% BSA in 0.1% Tween-20 in Tris-buffered saline containing (TBST)], and incubated overnight at 4°C with the following primary antibodies: rabbit anti-phosphorylated Stat1 (clone 58D6, 1:1000; #9167, Cell Signalling) or rabbit anti-Bdnf (1:1000; ab108319, Abcam).

Techniques: Comparison, Western Blot, Cell Culture, Staining

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: si-RNA sequences.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Sequencing, Negative Control

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: Sequences of primers used for reverse transcription-quantitative PCR.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Sequencing

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: STAT1 binds to the BST2 promoter to regulate BST2 expression in oral squamous cell carcinoma. (A) JASPAR and UCSC databases were used to predict the binding sites between STAT1 and the BST2 promoter. (B) mRNA expression levels of STAT1 and BST2 in SCC-15 cell lines were assessed using reverse transcription-quantitative PCR after regulation of STAT1 or BST2. (C) Relative luciferase activity was assessed using the dual-luciferase reporter assay in SCC-15 cells co-transfected with oe-STAT1 or oe-NC and BST2-WT or BST2-MUT. ** P<0.01 and *** P<0.001. BST2, bone marrow stromal antigen 2; oe, overexpression; WT, wildtype; MUT, mutant; NS, not significant; UCSC, University of California Santa Cruz Genome Browser.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Expressing, Binding Assay, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay, Reporter Assay, Transfection, Over Expression, Mutagenesis

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: STAT1 is highly expressed in HNSC and OSCC cell lines. (A) The mRNA expression levels of STAT1 from the Gene Expression Profiling Interactive Analysis database in different tumors. (B) Differential expression of STAT1 between normal and tumor tissues (tumor tissues, n=519; normal tissues, n=44). (C) Kyoto Encyclopedia of Genes and Genomes pathway analysis performed using the linkedomics database. (D) The protein and mRNA expression levels of STAT1 in different OSCC cell lines. * P<0.05 and *** P<0.001. HNSC, head and neck squamous cell carcinoma; OSCC, oral squamous cell carcinoma; TPM, transcripts per million; FDR, false discovery rate; NS, not significant; T, tumor; N, normal.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: STAT1 positively regulates BST2/AKT/ERK1/2 and biological behavior of OSCC. (A) Western blotting analysis of the protein expression levels of STAT1, BST2, AKT and ERK1/2 and the extent of AKT and ERK1/2 phosphorylation after using si-RNA or oe plasmid to regulate STAT1 or BST2 in SCC-15 cells. (B) Cell scratch test assay (magnification, ×50), and (C) Transwell assay (magnification, ×100) were used to assess the migration and invasion ability of SCC-15 cells after STAT1 downregulation. (D) Clone formation and (E) cell proliferation assays were performed to assess changes in the viability and proliferation of SCC-15 cells after STAT1 downregulation. * P<0.05, ** P<0.01 and *** P<0.001. BST2, bone marrow stromal antigen 2; si, small interfering; oe, overexpression; NC, negative control; p, phosphorylated; NS, not significant.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Western Blot, Expressing, Plasmid Preparation, Wound Healing Assay, Transwell Assay, Migration, Over Expression, Negative Control

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: STAT1 and BST2 serve a synergistic role in the biological behavior of oral squamous cell carcinoma. (A) Western blotting analysis of the protein expression levels of STAT1, BST2, AKT, ERK1/2 and the extent of AKT and ERK1/2 phosphorylation after using si-RNA or oe plasmid to regulate STAT1 or BST2 in SCC-15 cells. (B) Cell scratch test assay (magnification, ×50), (C) Transwell assay (magnification, ×100) and (D) clone formation assay were used to assess the migration, invasion and proliferation of SCC-15 cells after using si-RNA or oe plasmid to regulate STAT1 or BST2. * P<0.05, ** P<0.01 and *** P<0.001. BST2, bone marrow stromal antigen 2; si, small interfering; oe, overexpression; NC, negative control; p, phosphorylated; NS, not significant.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Western Blot, Expressing, Plasmid Preparation, Wound Healing Assay, Transwell Assay, Tube Formation Assay, Migration, Over Expression, Negative Control

Journal: International Journal of Oncology

Article Title: BST2 regulated by the transcription factor STAT1 can promote metastasis, invasion and proliferation of oral squamous cell carcinoma via the AKT/ERK1/2 signaling pathway

doi: 10.3892/ijo.2023.5502

Figure Lengend Snippet: (A) Photograph of tumors excised from mice, growth curve of subcutaneous tumor volume, and the weight of mice. (B) The protein expression levels of STAT1, BST2, AKT and ERK1/2 and the extent of STAT1, AKT and ERK1/2 phosphorylation in vivo after downregulation of STAT1 (n=3 selected from each group) and mRNA expression levels of STAT1 and BST2. Representative images of (C) hematoxylin and eosin and (D) immunohistochemical staining for STAT1, p-STAT1 and BST2 of oral squamous cell carcinoma tumor samples (magnification, ×200). * P<0.05, ** P<0.01 and *** P<0.001. BST2, bone marrow stromal antigen 2; si, small interfering; oe, overexpression; NC, negative control; p, phosphorylated; NS, not significant.

Article Snippet: The membranes were then incubated overnight with primary antibodies against STAT1 (1:1,000; cat. no. 10144-2-AP; ProteinTech Group, Inc.), phosphorylated (p)-STAT1 (1:1,000; cat. no. ab109461; Abcam), AKT (pan; 1:1,000; cat. no. 4691S; Cell Signaling Technology, Inc.), p-AKT (1:2,000; cat. no. 4060S; Cell Signaling Technology, Inc.), ERK1/2 (1:1,000; cat. no. 11257-1-AP; ProteinTech Group, Inc.), p-ERK1/2 (1:1,000; cat. no. 28733-1-AP-ProteinTech Group, Inc.) or β-actin (1:2,000; cat. no. 20536-1-AP; ProteinTech Group, Inc.) at 4°C.

Techniques: Expressing, In Vivo, Immunohistochemical staining, Staining, Over Expression, Negative Control

Journal: Scientific Reports

Article Title: Comparison of anti-inflammatory and anti-angiogenic effects of JAK inhibitors in IL-6 and TNFα-stimulated fibroblast-like synoviocytes derived from patients with RA

doi: 10.1038/s41598-025-94894-2

Figure Lengend Snippet: ( a ) Western blotting was performed to analyze STAT1, pSTAT1, STAT3, pSTAT3, and β-actin. ( b , c ) Comparison of the ratios of pSTAT1 to STAT1 and pSTAT3 to STAT1. Protein expression was determined by semi-quantitative analysis of digitally captured images. All JAK inhibitors significantly suppressed pSTAT1 and pSTAT3 levels compared to those in the control. Compared to TOF, BAR, PEF, UPA, and FIL suppressed pSTAT1 and pSTAT3 levels. STAT, signal transducer and activator of transcription; JAK, Janus kinase; TOF, tofacitinib; BAR, baricitinib; PEF, peficitinib; UPA, upadacitinib; FIL, filgotinib.

Article Snippet: The membrane was blocked with 5% skimmed milk in TBST at 25 °C for 30 min, incubated with antibodies against anti-STAT1, anti-phosphorylated STAT1 (pSTAT1), anti-STAT3, and anti-pSTAT3 (Cell Signaling Technology, Danvers, MA, US) at 4 °C for 12 h, and further incubated with horseradish peroxidase-conjugated goat anti-rabbit IgG secondary antibody at 25 °C for 1 h. The proteins were subsequently visualized using ECL Plus reagent (GE Healthcare Life Sciences, Little Chalfont, UK) on a chemiluminescence analyzer (LAS-3000 mini; Fujifilm, Tokyo, Japan).

Techniques: Western Blot, Comparison, Expressing, Control

Journal: Journal of Neuroinflammation

Article Title: A systems pharmacology-based approach to identify novel Kv1.3 channel-dependent mechanisms in microglial activation

doi: 10.1186/s12974-017-0906-6

Figure Lengend Snippet: Identification of Kv1.3-regulated signaling mechanisms in LPS-induced microglial activation. a , b Canonical pathway analysis of 120 Kv1.3-dependent proteins identified in our proteomic dataset revealed highly represented signaling pathways, two of which are shown here (see Additional file : Table S5 for others). Members of this list of 120 proteins are marked with ( red circle ). Transcription factors are also highlighted ( transparent red circle ). Arrows indicate directionality of the interaction (upstream vs. downstream). a GABPA, a Kv1.3-dependent transcription factor that was downregulated by LPS, was placed upstream of several Kv1.3-regulated proteins. b MHCI proteins of relevance to our results (TAP1, Tapasin, and EHD1 proteins) were represented in a signaling network that suggested that STAT1 and IRF1 may serve as upstream regulators of these proteins. c Comparison of normalized protein expression of transcription factors identified in our proteomic dataset across treatment groups. d Quantitative RT-PCR data comparing IRF1, IRF7, and NFKB1 mRNA expression across treatment groups (paired t tests were used for these comparisons; six replicates/group). e Phospho-flow cytometric studies of serine (S727) and tyrosine (Y701) STAT1 phosphorylation in BV2 microglia ( n = 5/treatment group) at 30-min and 3-h time points. f Immunofluorescence microscopy showing partial co-localization between Kv1.3 ( green , detected by ShK-F6CA labeling) and CD14 ( red ) in LPS-activated BV2 microglia (* p < 0.05, ** p < 0.01, *** p < 0.005)

Article Snippet: Phospho-flow cytometry was performed to detect pS727 and pY701 STAT1 phosphorylation (pY701-Alexa488 ab: Cell Signaling, pS727-Alexa647: BD Biosciences) [ ].

Techniques: Activation Assay, Protein-Protein interactions, Comparison, Expressing, Quantitative RT-PCR, Phospho-proteomics, Immunofluorescence, Microscopy, Labeling

Journal: Kidney & blood pressure research

Article Title: Cylindromatosis (CYLD), a Deubiquitinase, Attenuates Inflammatory Signaling Pathways by Activating Toll-Like Receptor 3 in Human Mesangial Cells.

doi: 10.1159/000485084

Figure Lengend Snippet: Fig. 2. Poly IC-induced expression of RIG-I and MDA5 mRNAs was decreased but the corresponding proteins were increased by CYLD silencing.Cells were transfected with siRNA against CYLD and incubated for 48 h as in Fig. 1. Cells were then treated with 30 μg/mL poly IC. (A) After 16 h incubation, RNA was extracted from the cells and quantitative real-time RT-PCR was performed (n=3, *p<0.01). (B) After incubation for 6 h (for STAT1 and p-STAT1) or 24 h (for RIG-I and MDA5), the cells were lysed and expression of CYLD, STAT1, p-STAT1, RIG-I, MDA5, and actin was determined by western blotting. The intensity of the bands was quantified using image J software, and normalized with actin.

Article Snippet: Mouse IgG against anti-phosphorylated STAT1 (signal transducers and activator of transcription protein 1) (p-STAT1) (sc-136229) and rabbit anti-STAT1 (sc-592) were from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Transfection, Incubation, Quantitative RT-PCR, Western Blot, Software