anti-chicken p-stat1 (ser 727)  (Santa Cruz Biotechnology)

Journal: Scientific reports

Article Title: Inhibitory effect of Sanguisorba hakusanensis Makino ethanol extract on atopic dermatitis-like responses in NC/Nga mice and human keratinocytes.

doi: 10.1038/s41598-023-41676-3

Figure Lengend Snippet: Figure 5. SHE inhibits the MAPK/NF-κB/STAT1 signaling pathway in IFN-γ/TNF-α-stimulated HaCaT cells. The cells were pre-treated with SHE (300 µg/mL) for 1 h and then stimulated with IFN-γ/TNF-α for 0, 5, 15, and 30 min. (a) Phosphorylated and total MAPK (p38, ERK, and JNK) protein levels as well as (b) phosphorylation or degradation of IκBα and STAT1 were detected in the cells. (c, d) HaCaT cells were pre- stimulated with SHE for 1 h and then treated with IFN-γ/TNF-α for 1 h. (c) Nuclear extracts were analyzed using western blotting to detect p-STAT1, STAT1, and NF-κB (p65 and p50 subunits). PCNA was used as a loading control for nuclear extracts. For clarity of data, the Western blots were cropped and all original blots are presented in Supplementary Figure 1. (d) Localization of p-STAT1 (green and red) was visualized using immunofluorescence. The nuclei were stained with DRAQ (blue). Scale bar = 10 μm.

Article Snippet: Phospho (p)-STAT1 (Tyr), p-STAT1 (Ser), STAT1, p-IκBα, IκBα, p-ERK, ERK, p-p38, p38, p-JNK, JNK, p65, and β-actin antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Phospho-proteomics, Western Blot, Control, Immunofluorescence, Staining

Journal: Immunologic Research

Article Title: Tryptanthrin attenuates TLR3-mediated STAT1 activation in THP-1 cells

doi: 10.1007/s12026-022-09301-z

Figure Lengend Snippet: A THP-1 Mϕ were stimulated with 20 μg/mL polyinosinic-polycytidylic acid (poly IC) for 8 h. The cell lysates were subjected to western blotting to analyze the expression of p-STAT1-Ser, p-STAT1-Tyr, and STAT1. B Cells were stimulated using 1 μg/mL lipopolysaccharides (LPS) for 4 h. The cell lysates were subjected to western blotting to analyze the expression of p-STAT1-Ser, p-STAT1-Tyr, and STAT1. C Cells were stimulated with 20 μg/mL poly IC and 0.01–1 μM tryptanthrin for 8 h. The expression of p-STAT1-Ser, p-STAT1-Tyr, and STAT1 proteins was analyzed using western blotting. Cells were stimulated with 1 μg/mL LPS and 0.01–1 μM tryptanthrin, and the expression of p-STAT1-Ser ( D ) and p-STAT1-Tyr ( E ) was analyzed at 1 h and 4 h, respectively. Representative images of three independent experiments

Article Snippet: Rabbit monoclonal anti-phosphorylated IRF3 (p-IRF3) antibody (#4947) and rabbit polyclonal anti-phosphorylated STAT1 (Ser727) (p-STAT1-Ser) antibody (#9177) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Expressing

Journal: Immunologic Research

Article Title: Tryptanthrin attenuates TLR3-mediated STAT1 activation in THP-1 cells

doi: 10.1007/s12026-022-09301-z

Figure Lengend Snippet: A THP-1 Mϕ were stimulated with 20 μg/mL poly IC or 1 μg/mL LPS for 8 h or 4 h, respectively, with or without tryptanthrin. The cells were fixed, and then immunostained with anti-STAT1 antibody (red). Cell nuclei were stained with DAPI (blue). Yellow arrowhead indicates the nuclear translocation of STAT1. Representative images of three independent experiments. B THP-1 Mϕ were stimulated with 20 μg/mL poly IC or vehicle (control), and 1 μM tryptanthrin or DMSO for 8 h. Thereafter, the total RNA was extracted from the cells and expression of IFIT1 , IFIT2 , IFITM1 , ISG15 , and Mx1 mRNA was analyzed using qRT-PCR. C Similarly, after stimulating the cells with 1 μg/mL LPS or vehicle, and 1 μM tryptanthrin or DMSO for 4 h, total RNA was extracted and the mRNA expression was examined ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001; n.s., not significant)

Article Snippet: Rabbit monoclonal anti-phosphorylated IRF3 (p-IRF3) antibody (#4947) and rabbit polyclonal anti-phosphorylated STAT1 (Ser727) (p-STAT1-Ser) antibody (#9177) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Staining, Translocation Assay, Control, Expressing, Quantitative RT-PCR

Journal: Immunologic Research

Article Title: Tryptanthrin attenuates TLR3-mediated STAT1 activation in THP-1 cells

doi: 10.1007/s12026-022-09301-z

Figure Lengend Snippet: A Cells were incubated with 1 μM tryptanthrin or DMSO for 24 h, and the total RNA was extracted. The mRNA expression of toll-like receptor 3 (TLR3) was examined by qRT-PCR ( n = 3; n.s., not significant). B Flow cytometry analysis of TLR3 expression in THP-1 Mϕ treated with 1 μM tryptanthrin (right panel) or DMSO (left panel). Representative histograms of three to five independent experiments. C Quantification of mean fluorescence intensity for TLR3 ( n = 3; n.s., not significant). D Cells were incubated with 1 μM tryptanthrin or DMSO for 24 h, fixed with methanol, and immunostained with anti-TLR3 antibody (green). Cell nuclei were stained using DAPI (blue). Representative images of three independent experiments. E , F Cells were stimulated with 1 ng/mL rIFN-β ( E ) or rIFN-γ ( F ) with or without tryptanthrin for up to 30 min, and the cell lysates were subjected to western blotting to analyze p-STAT1-Tyr and STAT1. G Putative mechanism of tryptanthrin-mediated regulation of TLR3 signaling in THP Mϕ. Tryptanthrin negatively modulates IRF3 activation and the subsequent IFN-β-induced phosphorylation of STAT1

Article Snippet: Rabbit monoclonal anti-phosphorylated IRF3 (p-IRF3) antibody (#4947) and rabbit polyclonal anti-phosphorylated STAT1 (Ser727) (p-STAT1-Ser) antibody (#9177) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Incubation, Expressing, Quantitative RT-PCR, Flow Cytometry, Fluorescence, Staining, Western Blot, Activation Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-34 Regulates Th1 and Th17 Cytokine Production by Activating Multiple Signaling Pathways through CSF-1R in Chicken Cell Lines

doi: 10.3390/ijms19061665

Figure Lengend Snippet: Comparison the function of interleukin (IL)-34 in mammals and chickens.

Article Snippet: The rabbit anti-chicken p-STAT1 (Ser 727 ), rabbit anti-chicken p-STAT3 (Ser 727 ), and rabbit anti-chicken p-JAK2 (Tyr 1007 /Tyr 1008 ) antibodies were purchased from Santa Cruz Biotech (Dallas, TX, USA).

Techniques: Marker

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-34 Regulates Th1 and Th17 Cytokine Production by Activating Multiple Signaling Pathways through CSF-1R in Chicken Cell Lines

doi: 10.3390/ijms19061665

Figure Lengend Snippet: Primer sequences for qRT-PCR analyses of gene expression levels.

Article Snippet: The rabbit anti-chicken p-STAT1 (Ser 727 ), rabbit anti-chicken p-STAT3 (Ser 727 ), and rabbit anti-chicken p-JAK2 (Tyr 1007 /Tyr 1008 ) antibodies were purchased from Santa Cruz Biotech (Dallas, TX, USA).

Techniques: Expressing, Sequencing