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Cell Signaling Technology Inc rabbit phosphor stat1 y701 antibody cst
Rabbit Phosphor Stat1 Y701 Antibody Cst, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit phosphor stat1 y701 antibody cst/product/Cell Signaling Technology Inc
Average 96 stars, based on 225 article reviews

rabbit phosphor stat1 y701 antibody cst - by Bioz Stars, 2026-03

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Suppression of STAT1 and STAT2 phosphorylation by NSs. A , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells transfected with expression plasmid for HA-tagged NSs were treated with IFN-αA/D (2000 U/ml) or left untreated for 30 min and were then lysed for detection of each protein expression by immunoblotting. B and C , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells were infected with SFTSV at an MOI of 10. C , after 18 h or 48 h, these cells were treated with IFN-αA/D (2000 U/ml) or left untreated for 30 min and were then lysed for detection of each protein expression by immunoblotting. The relative expression rates of pSTAT1 and pSTAT2 in mock-transfected cells and uninfected cells treated with IFN-I are set at 100%. The assays were independently performed in triplicate. The data represent averages with SDs. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFN, interferon; MOI, multiplicity of infection; NSs, nonstructural protein; SFTSV, severe fever with thrombocytopenia syndrome virus; STAT, signal transducer and activator of transcription.

Journal: The Journal of Biological Chemistry

Article Title: The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species

doi: 10.1016/j.jbc.2023.104819

Figure Lengend Snippet: Suppression of STAT1 and STAT2 phosphorylation by NSs. A , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells transfected with expression plasmid for HA-tagged NSs were treated with IFN-αA/D (2000 U/ml) or left untreated for 30 min and were then lysed for detection of each protein expression by immunoblotting. B and C , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells were infected with SFTSV at an MOI of 10. C , after 18 h or 48 h, these cells were treated with IFN-αA/D (2000 U/ml) or left untreated for 30 min and were then lysed for detection of each protein expression by immunoblotting. The relative expression rates of pSTAT1 and pSTAT2 in mock-transfected cells and uninfected cells treated with IFN-I are set at 100%. The assays were independently performed in triplicate. The data represent averages with SDs. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFN, interferon; MOI, multiplicity of infection; NSs, nonstructural protein; SFTSV, severe fever with thrombocytopenia syndrome virus; STAT, signal transducer and activator of transcription.

Article Snippet: Western blotting was performed as previously described using the following antibodies ( , ): anti-HA (catalog no.: 18850; QED Biosciences, Inc), anti-His (catalog no.: 9F2; Wako), anti-FLAG (catalog no.: F7425; Sigma–Aldrich), anti-STAT2 (catalog no.: D9J7L; Cell Signaling Technology), anti-STAT1 (catalog no.: D19KY; Cell Signaling Technology), anti-STAT2 (phosphor Y690) (catalog no.: GTX50721; GeneTex), anti-STAT1 (phosphor Y701) (catalog no.: D4A7; Cell Signaling Technology), or anti-β-actin (catalog no.: AC-15; Sigma–Aldrich).

Techniques: Transfection, Expressing, Plasmid Preparation, Western Blot, Infection

Influence of NSs to STAT1 and STAT2 activation in cells derived from different animal species. HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells were transfected with the expression plasmid for HA-tagged NSs and treated with IFN-αA/D (2000 U/ml) for 30 min at 48 h post-transfection. IFA was performed to detect NSs, the nuclei, and pSTAT1 or pSTAT2, shown in red , blue , and green , respectively. Each figure in the red frame and blue frame indicates the result of indirect IFA with anti-pSTAT1 and anti-pSTAT2, respectively. The a rrow and arrowhead indicate NSs-expressing and nonexpressing cells, respectively. Scale bar represents 50 μm. Representative results of IFA are shown. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFA, immunofluorescence assay; IFN, interferon; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Journal: The Journal of Biological Chemistry

Article Title: The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species

doi: 10.1016/j.jbc.2023.104819

Figure Lengend Snippet: Influence of NSs to STAT1 and STAT2 activation in cells derived from different animal species. HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), or PK15 (pig) cells were transfected with the expression plasmid for HA-tagged NSs and treated with IFN-αA/D (2000 U/ml) for 30 min at 48 h post-transfection. IFA was performed to detect NSs, the nuclei, and pSTAT1 or pSTAT2, shown in red , blue , and green , respectively. Each figure in the red frame and blue frame indicates the result of indirect IFA with anti-pSTAT1 and anti-pSTAT2, respectively. The a rrow and arrowhead indicate NSs-expressing and nonexpressing cells, respectively. Scale bar represents 50 μm. Representative results of IFA are shown. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFA, immunofluorescence assay; IFN, interferon; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Techniques: Activation Assay, Derivative Assay, Transfection, Expressing, Plasmid Preparation, Immunofluorescence

Interaction of NSs with STAT2. A , NIH3T3 (mouse) cells were transfected with the expression plasmid for HA-tagged NSs and each of the His-tagged STAT2, and then co-IP assays were performed. B , colocalization of NSs with STAT2. NIH3T3 (mouse) cells were transfected with the expression plasmid for HA-tagged NSs and each of the His-tagged STAT2. IFA was also performed with NSs, STAT2, and the nuclei, shown in green , red , and blue , respectively. Scale bar represents 20 μm. C , identification of binding regions in porcine STAT2 to NSs. Schematic representation of the chimeric mutants of human, mouse, and pig STAT2 ( upper ). NIH3T3 (mouse) cells were transfected with the HA-tagged NSs and each of the His-tagged STAT2 chimeras, following which co-IP assays were performed ( lower ). D , the phylogenetic tree of the 101 to 315 region of each STAT2. This phylogenetic tree was constructed using the neighbor-joining method. The bootstrap values are indicated on each node. Representative results of Western blotting assays ( A and C ) and IFA ( B ) are shown. co-IP, coimmunoprecipitation; HA, hemagglutinin; IFA, immunofluorescence assay; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Journal: The Journal of Biological Chemistry

Article Title: The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species

doi: 10.1016/j.jbc.2023.104819

Figure Lengend Snippet: Interaction of NSs with STAT2. A , NIH3T3 (mouse) cells were transfected with the expression plasmid for HA-tagged NSs and each of the His-tagged STAT2, and then co-IP assays were performed. B , colocalization of NSs with STAT2. NIH3T3 (mouse) cells were transfected with the expression plasmid for HA-tagged NSs and each of the His-tagged STAT2. IFA was also performed with NSs, STAT2, and the nuclei, shown in green , red , and blue , respectively. Scale bar represents 20 μm. C , identification of binding regions in porcine STAT2 to NSs. Schematic representation of the chimeric mutants of human, mouse, and pig STAT2 ( upper ). NIH3T3 (mouse) cells were transfected with the HA-tagged NSs and each of the His-tagged STAT2 chimeras, following which co-IP assays were performed ( lower ). D , the phylogenetic tree of the 101 to 315 region of each STAT2. This phylogenetic tree was constructed using the neighbor-joining method. The bootstrap values are indicated on each node. Representative results of Western blotting assays ( A and C ) and IFA ( B ) are shown. co-IP, coimmunoprecipitation; HA, hemagglutinin; IFA, immunofluorescence assay; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Techniques: Transfection, Expressing, Plasmid Preparation, Co-Immunoprecipitation Assay, Binding Assay, Construct, Western Blot, Immunofluorescence

Function of human, murine, porcine, and chimeric STAT2s in SFTSV infection. SFTSV was inoculated into HEK293T (human) cells transfected with expression plasmid for each STAT2 at MOI 1. After 24 hpi, cells were treated with IFN-αA/D (500 U/ml) for 48 h and then collected culture supernatants. The SFTSV titer in culture supernatants was determined by focus-forming assay ( upper ). The protein expression was detected by Western blotting ( lower ). The assays were independently performed in triplicate. Values are the averages with SDs of data from nine results obtained from three experiments (n = 9). ∗∗ p < 0.01, versus human STAT2. Each exact p value, average, and SD is shown in <xref ref-type=

Table S4 . HEK293T, human embryonic kidney 293T cell line; hpi, hours postinfection; IFN, interferon; MOI, multiplicity of infection; SFTSV, severe fever with thrombocytopenia syndrome virus; STAT, signal transducer and activator of transcription. " width="100%" height="100%">

Journal: The Journal of Biological Chemistry

Article Title: The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species

doi: 10.1016/j.jbc.2023.104819

Figure Lengend Snippet: Function of human, murine, porcine, and chimeric STAT2s in SFTSV infection. SFTSV was inoculated into HEK293T (human) cells transfected with expression plasmid for each STAT2 at MOI 1. After 24 hpi, cells were treated with IFN-αA/D (500 U/ml) for 48 h and then collected culture supernatants. The SFTSV titer in culture supernatants was determined by focus-forming assay ( upper ). The protein expression was detected by Western blotting ( lower ). The assays were independently performed in triplicate. Values are the averages with SDs of data from nine results obtained from three experiments (n = 9). ∗∗ p < 0.01, versus human STAT2. Each exact p value, average, and SD is shown in Table S4 . HEK293T, human embryonic kidney 293T cell line; hpi, hours postinfection; IFN, interferon; MOI, multiplicity of infection; SFTSV, severe fever with thrombocytopenia syndrome virus; STAT, signal transducer and activator of transcription.

Techniques: Infection, Transfection, Expressing, Plasmid Preparation, Focus Forming Assay, Western Blot

Interaction of NSs with STAT1. A , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), and PK15 (pig) cells were transfected with the expression plasmid for HA-tagged NSs, and then co-IP assays were performed. The relative binding rates of NSs to human STAT1 are set at 100%. The assays were independently performed in triplicate. The data represent averages with SDs. The asterisks indicate the nonspecific bands. B , colocalization of NSs with STAT2. The expression plasmid for HA-tagged NSs was transfected to HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), and PK15 (pig) cells. IFA was also performed with NSs, STAT1, and the nuclei, shown in green , red , and blue , respectively. Scale bar represents 20 μm. C , interaction of NSs with exogenous STAT1. HEK293T (human), NIH3T3 (mouse), and CRFK (cat) cells were transfected with the expression plasmid for HA-tagged NSs, His-tagged human STAT1 (hSTAT1), murine STAT1 (mSTAT1), feline STAT1 (fSTAT1), and human STAT2 (hSTAT2), and then co-IP assays were performed. D , Interaction activity of NSs to STAT1 and STAT2. The expression plasmid for HA-tagged NSs was transfected into NIH3T3 (mouse) cells with the expression plasmid for His-tagged human STAT1(H) or feline STAT1(F) and FLAG-tagged human STAT2 or feline STAT2, and then co-IP assays were performed. Representative results of Western blotting assays ( A , C , and D ) and IFA ( B ) are shown. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFA, immunofluorescence assay; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Journal: The Journal of Biological Chemistry

Article Title: The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species

doi: 10.1016/j.jbc.2023.104819

Figure Lengend Snippet: Interaction of NSs with STAT1. A , HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), and PK15 (pig) cells were transfected with the expression plasmid for HA-tagged NSs, and then co-IP assays were performed. The relative binding rates of NSs to human STAT1 are set at 100%. The assays were independently performed in triplicate. The data represent averages with SDs. The asterisks indicate the nonspecific bands. B , colocalization of NSs with STAT2. The expression plasmid for HA-tagged NSs was transfected to HEK293T (human), NIH3T3 (mouse), CRFK (cat), A72 (dog), Mpf (ferret), and PK15 (pig) cells. IFA was also performed with NSs, STAT1, and the nuclei, shown in green , red , and blue , respectively. Scale bar represents 20 μm. C , interaction of NSs with exogenous STAT1. HEK293T (human), NIH3T3 (mouse), and CRFK (cat) cells were transfected with the expression plasmid for HA-tagged NSs, His-tagged human STAT1 (hSTAT1), murine STAT1 (mSTAT1), feline STAT1 (fSTAT1), and human STAT2 (hSTAT2), and then co-IP assays were performed. D , Interaction activity of NSs to STAT1 and STAT2. The expression plasmid for HA-tagged NSs was transfected into NIH3T3 (mouse) cells with the expression plasmid for His-tagged human STAT1(H) or feline STAT1(F) and FLAG-tagged human STAT2 or feline STAT2, and then co-IP assays were performed. Representative results of Western blotting assays ( A , C , and D ) and IFA ( B ) are shown. CRFK, Crandell–Rees feline kidney; HA, hemagglutinin; HEK293T, human embryonic kidney 293T cell line; IFA, immunofluorescence assay; NSs, nonstructural protein; STAT, signal transducer and activator of transcription.

Techniques: Transfection, Expressing, Plasmid Preparation, Co-Immunoprecipitation Assay, Binding Assay, Activity Assay, Western Blot, Immunofluorescence

FIGURE 2 ASFV pA104R inhibits ISGF3-induced ISRE promoter activity and attenuates the phosphorylation of STAT1. (A) HEK-293 T cells were transfected with various concentrations of pA104R plasmids, along with ISGF3 complex (STAT1, STAT2 and IRF9) and pISRE-Luc and pRL- TK plasmids. After 30 h, a luciferase assay was performed. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. (B) HEK-293 T cells were transfected with pA104R or empty vector. After 24 h, cells were treated with 1,000 U/ ml IFN-α for 2 h. The levels of total or phosphorylated STAT1, STAT2, and IRF9 were detected by immunoblotting analysis.

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 2 ASFV pA104R inhibits ISGF3-induced ISRE promoter activity and attenuates the phosphorylation of STAT1. (A) HEK-293 T cells were transfected with various concentrations of pA104R plasmids, along with ISGF3 complex (STAT1, STAT2 and IRF9) and pISRE-Luc and pRL- TK plasmids. After 30 h, a luciferase assay was performed. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. (B) HEK-293 T cells were transfected with pA104R or empty vector. After 24 h, cells were treated with 1,000 U/ ml IFN-α for 2 h. The levels of total or phosphorylated STAT1, STAT2, and IRF9 were detected by immunoblotting analysis.

Article Snippet: The STAT1 (9172), STAT2 (72604), phosphor-STAT1 (9649), phosphor-STAT2 (88410), and IRF9 (76684) antibodies were purchased from Cell Signaling Technology (Danvers, MA). β-Actin (66009-1-Ig), GFP-tag (66002-1-Ig), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP) antibodies were purchased from Proteintech (Chicago, IL).

Techniques: Activity Assay, Phospho-proteomics, Transfection, Luciferase, Plasmid Preparation, Western Blot

FIGURE 3 ASFV pA104R does not interact with STAT1, STAT2, and IRF9. HEK-293 T cells were transfected with pA104R alone (C) or co-transfected with STAT1, STAT2, and IRF9 (A,B). After 24 h, cells were treated with 1,000 U/mL IFN-α for 8 h. Cell lysates were prepared and subjected to immunoprecipitation analysis. The whole-cell lysates and immunoprecipitation complexes were analyzed by immunoblotting with the indicated antibodies.

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 3 ASFV pA104R does not interact with STAT1, STAT2, and IRF9. HEK-293 T cells were transfected with pA104R alone (C) or co-transfected with STAT1, STAT2, and IRF9 (A,B). After 24 h, cells were treated with 1,000 U/mL IFN-α for 8 h. Cell lysates were prepared and subjected to immunoprecipitation analysis. The whole-cell lysates and immunoprecipitation complexes were analyzed by immunoblotting with the indicated antibodies.

Techniques: Transfection, Immunoprecipitation, Western Blot

FIGURE 5 ASFV pA104R does not prevent the association of ISGF3 with promoter. (A) The ISRE DNA and control oligonucleotides used for DNA pull-down. (B) HEK-293T cells were co-transfected with ISGF3 complex (STAT1, STAT2, and IRF9), along with pA104R or empty control. After 24 h post- transfection, cells were treated with 1,000 U/mL IFN-α for 12 h. Nuclear extracts were incubated with a Biotin-labeled ISRE or control probe and subjected to pull-down analysis with streptavidin magnetic beads. The whole-cell lysates and pull-down complexes were analyzed by immunoblotting with the indicated antibodies.

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 5 ASFV pA104R does not prevent the association of ISGF3 with promoter. (A) The ISRE DNA and control oligonucleotides used for DNA pull-down. (B) HEK-293T cells were co-transfected with ISGF3 complex (STAT1, STAT2, and IRF9), along with pA104R or empty control. After 24 h post- transfection, cells were treated with 1,000 U/mL IFN-α for 12 h. Nuclear extracts were incubated with a Biotin-labeled ISRE or control probe and subjected to pull-down analysis with streptavidin magnetic beads. The whole-cell lysates and pull-down complexes were analyzed by immunoblotting with the indicated antibodies.

Techniques: Control, Transfection, Incubation, Labeling, Magnetic Beads, Western Blot

FIGURE 6 ASFV pA104R does not exert repressive effects on trans-activation domains and transcriptional co-stimulatory factors. (A) HEK-293 T cells were co- transfected with pA104R and IRF9-Stat2(TA) (A) or GAL4-Stat2(TA) (B), along with pRL-TK and pISRE-Luc (A) or pGAL4-UAS-Luc (B) plasmids. After 30 h, a luciferase assay was performed. (C) HEK-293 T Cells were co-transfected with pA104R and CBP or p300 along with pISRE-Luc and pRL-TK plasmids. After 24 h post-transfection, cells were treated with 1,000 U/mL IFN-α for 12 h, followed by luciferase assays. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. *** p < 0.001.

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 6 ASFV pA104R does not exert repressive effects on trans-activation domains and transcriptional co-stimulatory factors. (A) HEK-293 T cells were co- transfected with pA104R and IRF9-Stat2(TA) (A) or GAL4-Stat2(TA) (B), along with pRL-TK and pISRE-Luc (A) or pGAL4-UAS-Luc (B) plasmids. After 30 h, a luciferase assay was performed. (C) HEK-293 T Cells were co-transfected with pA104R and CBP or p300 along with pISRE-Luc and pRL-TK plasmids. After 24 h post-transfection, cells were treated with 1,000 U/mL IFN-α for 12 h, followed by luciferase assays. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. *** p < 0.001.

Techniques: Activation Assay, Transfection, Luciferase

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