rabbit anti-stat1 monoclonal antibodies (mab) (14994)  (Cell Signaling Technology Inc)

Journal: Cell death discovery

Article Title: TBK1 is involved in programmed cell death and ALS-related pathways in novel zebrafish models.

doi: 10.1038/s41420-025-02374-3

Figure Lengend Snippet: Fig. 6 Joint proteomic analysis of tbk1 mutants zebrafish and iMNs from ALS patients with TBK1 mutations. A Fluorescent immunolabeling of the neuronal marker CHAT in differentiated induced pluripotent stem cell-derived motor neurons (iMNs) at day 21 from an ALS patient harboring the loss of function mutation TBK1T77fs compared to control iMNs. B Proteomic profiles of ALS-TBK1 iMNs compared to control iMNs. The differentially expressed protein list was compared to the list of zebrafish orthologs differentially expressed in tbk1-deficient zebrafish larvae, and enrichment analysis was performed on the common list of 260 terms. C Network plot showing the relationship between enriched pathways from TBK1-mutant ALS models. A close interaction between the necroptosis pathway and several neurodegenerative pathways, including the amyotrophic lateral sclerosis (ALS) KEGG pathway, is observed. D Heatmap representation of protein levels (row z scores) for significantly enriched proteins in both datasets associated with the KEGG term ALS in tbk1 mutant vs NI zebrafish and ALS-TBK1 iMNs vs control iMNs. E Heatmap representation of protein levels (row z scores) for significantly enriched proteins associated with the KEGG term necroptosis in ALS-TBK1 iMNs vs control iMNs. F Western blotting of STAT1 expression in tbk1 mutant and NI zebrafish. G Mass spectrometry quantification of stat1b protein level in tbk1 mutant and NI zebrafish.

Article Snippet: The separated proteins were transferred to nitrocellulose membranes (0.45 μm, Life Sciences) and probed with the following primary antibody: NAK1/TBK1 antibody (ab109735, Abcam), STAT1 (D1K9Y) antibody (#14994 Cell signaling) and α-tubulin antibody (T5168, Sigma‒ Aldrich) was used as a loading control.

Techniques: Immunolabeling, Marker, Derivative Assay, Mutagenesis, Control, Western Blot, Expressing, Mass Spectrometry

Journal: Communications Biology

Article Title: STAT1 mediates the pro-inflammatory role of GBP5 in colitis

doi: 10.1038/s42003-025-07843-0

Figure Lengend Snippet: a Plots are the mRNA expression data of STAT1 and STAT1 target genes from RNA sequencing-based transcriptome analysis. Wildtype and GBP5 -/- THP-1 cells were treated with IFNγ (25 ng/ml) plus LPS (500 ng/ml) or mock-treated for 16 h. The list of the STAT1 target genes was from TRRUST version 2, with a few others ( CCND1 , MYC , and CDC25A ) added according to the literature. Genes known to be stimulated and repressed by STAT1 are indicated by red and green, respectively. n = 3 biologically independent samples for each group. b Gene set enrichment analysis (GSEA) of the entire transcriptome data of GBP5 -/- cells compared to wild-type THP-1 cells. The upper panel is the enrichment score curve for JAK-STAT signaling pathway, showing the decreased expression level of relevant genes in Gbp5 deficient cells. The middle panel shows the distribution of the genes related to JAK-STAT signaling pathway. The genes were ranked according to their differential expression between wild-type and GBP5 -/- cells. The lower panel is a graphical representation of the correlations between gene expression levels and the genotypes: WT or GBP5 -/- . NES, normalized enrichment scores; FDR, false discovery rate.

Article Snippet: Then, the rest of the protocol was followed by the instructions of immunohistochemistry (IHC) kit (ZSGB-BIO, Cat#PV-6000), with the GBP5 (1:200, CST, Cat#67798S) and STAT1 antibodies (1:200, CST, Cat#14994S).

Techniques: Expressing, RNA Sequencing, Quantitative Proteomics, Gene Expression

Journal: Communications Biology

Article Title: STAT1 mediates the pro-inflammatory role of GBP5 in colitis

doi: 10.1038/s42003-025-07843-0

Figure Lengend Snippet: a Expression of GBP5 and STAT1 in the wildtype and GBP5 -/- (three different clones A1, B1, and B2) THP-1 cells. THP-1 cells were treated with IFNγ (25 ng/ml), or IFNγ (25 ng/ml) plus LPS (500 ng/ml), or mock treated for 16 h. Cells were then harvested for Western blot analyses with anti-GBP5, anti-STAT1, and anti-β-actin antibodies, respectively. b Expression of GBP5 and STAT1 after siRNA knockdown of GBP5 in Jurkat cells. Jurkat cells were treated with GBP5 siRNA or control RNA for 48 h and subsequently stimulated with IFNγ/LPS for an additional 16 h. Cells were then harvested and subjected to Western blot analyses with anti-GBP5, anti-STAT1, and anti-GAPDH antibodies, respectively. c GBP5 -/- THP-1 cells were transfected with either control (CTRL), GBP5 or STAT1 plasmid for 48 h, followed by treatment with IFNγ/LPS for 16 h. The cells were then harvested and subjected to RT-qPCR analysis. n = 3 biologically independent samples for each group. Data are mean ± SEM, Student t -test. d GBP5 -/- THP-1 cells were transfected with either control (CTRL) or GBP5 plasmid for 48 h, followed by treatment with IFNγ/LPS for 16 h. Cells were then harvested and analyzed by Western blot with anti-Flag, anti-GBP5, anti-STAT1, anti-p-STAT1 (S727), and anti-β-actin antibodies, respectively. e Persistent expression of GBP5 despite the diminished expression of STAT1. THP-1 cells were treated with fludarabine (10 ng/ml) or mock-treated for 4 h, followed by IFNγ stimulation for 16 h. Cells were then harvested and analyzed by Western blot with anti-STAT1, anti-GBP5, and anti-β-actin antibodies, respectively. f UCSC genome browser tracks showing ChIP-seq of GBP5 (with or without IFNγ/LPS stimulation) and STAT1 (with or without IFNγ/LPS stimulation) bound promoter region of selected STAT1 effector genes in THP-1 cells.

Article Snippet: Then, the rest of the protocol was followed by the instructions of immunohistochemistry (IHC) kit (ZSGB-BIO, Cat#PV-6000), with the GBP5 (1:200, CST, Cat#67798S) and STAT1 antibodies (1:200, CST, Cat#14994S).

Techniques: Expressing, Clone Assay, Western Blot, Knockdown, Control, Transfection, Plasmid Preparation, Quantitative RT-PCR, ChIP-sequencing

Journal: Communications Biology

Article Title: STAT1 mediates the pro-inflammatory role of GBP5 in colitis

doi: 10.1038/s42003-025-07843-0

Figure Lengend Snippet: a Identification of STAT1 in the bound fraction of anti-GBP5 immunoprecipitation by liquid chromatograph followed by tandem mass spectrometry (LC-MS/MS). Wildtype THP-1 cells were treated with IFNγ/LPS or mock treated for 16 h. Cell lysates were immunoprecipitated with anti-GBP5. The bound fraction of the IFNγ/LPS treated sample was in-gel digested before LC-MS/MS analysis of the peptides. The collision-induced dissociation spectra of three STAT1 peptides are shown. b Western blot analyses of the total lysates (input) and bound fractions from immunoprecipitation with anti-GBP5. Lysates were prepared from wild-type THP-1 cells treated with IFNγ/LPS or mock-treated for 16 h. Blots were probed for GBP5, STAT1, and β-actin as indicated. c Western blot analyses of the total lysates (input) and bound fractions from immunoprecipitation with anti-STAT1, or IgG (control). Lysates were prepared from wild-type THP-1 cells treated with IFNγ/LPS or mock-treated for 16 h. Blots were probed for GBP5, STAT1, and β-actin as indicated. d Dock simulation for GBP5 and STAT1 interaction. The left panel represents the surface diagram of the docking model, where blue and yellow indicate GBP5 and STAT1, respectively. The right panel displays the interface of the docking model, highlighting the amino acid residues involved in the interaction.

Article Snippet: Then, the rest of the protocol was followed by the instructions of immunohistochemistry (IHC) kit (ZSGB-BIO, Cat#PV-6000), with the GBP5 (1:200, CST, Cat#67798S) and STAT1 antibodies (1:200, CST, Cat#14994S).

Techniques: Immunoprecipitation, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Western Blot, Control

Journal: Communications Biology

Article Title: STAT1 mediates the pro-inflammatory role of GBP5 in colitis

doi: 10.1038/s42003-025-07843-0

Figure Lengend Snippet: a Immunofluorescence staining of THP-1 cells untreated or treated with IFNγ/LPS using antibodies against GBP5 and STAT1. Scale bar: 5 μm. b Subcellular fractionation analysis of GBP5 and STAT1. WT and GBP5 -/- THP-1 cells were untreated or treated with IFNγ/LPS for 16 h, harvested to prepare nuclear and cytoplasmic fractions, and analyzed by Western blot with antibodies against GBP5, STAT1, and p-STAT1 (S727). GAPDH and Lamin B were used as internal references. The bar plot on the right was the quantification of three independent experiments. n = 3 biologically independent experiments for each group. Data are mean ± SEM, paired student t -test. c GBP5-STAT1 proximity ligation assay with THP-1 cells untreated or treated with IFNγ/LPS. Each red spot represents an interaction. Scale bar: 2 μm. Nuclei were stained with DAPI (blue).

Article Snippet: Then, the rest of the protocol was followed by the instructions of immunohistochemistry (IHC) kit (ZSGB-BIO, Cat#PV-6000), with the GBP5 (1:200, CST, Cat#67798S) and STAT1 antibodies (1:200, CST, Cat#14994S).

Techniques: Immunofluorescence, Staining, Fractionation, Western Blot, Proximity Ligation Assay