polyclonal rabbit anti-pstat2(y689) Search Results


polyclonal rabbit anti-pstat2(y689)  (Millipore)
90
Millipore polyclonal rabbit anti-pstat2(y689)
Polyclonal Rabbit Anti Pstat2(Y689), supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse polyclonal mx1  (Abnova)
90
Abnova mouse polyclonal mx1
Mouse Polyclonal Mx1, supplied by Abnova, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal ifit3  (Covance)
90
Covance rabbit polyclonal ifit3
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and <t>IFIT3,</t> and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Rabbit Polyclonal Ifit3, supplied by Covance, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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monoclonal mouse anti-irf-9(isgf3γ  (Becton Dickinson)
90
Becton Dickinson monoclonal mouse anti-irf-9(isgf3γ
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and <t>IFIT3,</t> and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Monoclonal Mouse Anti Irf 9(Isgf3γ, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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monoclonal mouse anti-pstat1(y701  (Becton Dickinson)
90
Becton Dickinson monoclonal mouse anti-pstat1(y701
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for <t>pSTAT1(Y701),</t> STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Monoclonal Mouse Anti Pstat1(Y701, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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monoclonal mouse anti actin  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc monoclonal mouse anti actin
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for <t>pSTAT1(Y701),</t> STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Monoclonal Mouse Anti Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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monoclonal anti irf 3  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc monoclonal anti irf 3
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for <t>pSTAT1(Y701),</t> STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Monoclonal Anti Irf 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pstat3  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc pstat3
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for <t>pSTAT1(Y701),</t> STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Pstat3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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p jak1 y1022 1023  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc p jak1 y1022 1023
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for <t>pSTAT1(Y701),</t> STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
P Jak1 Y1022 1023, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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polyclonal rabbit anti β tubulin  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc polyclonal rabbit anti β tubulin
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and <t>β-tubulin</t> were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Polyclonal Rabbit Anti β Tubulin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pstat3 s727 pstat1  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc pstat3 s727 pstat1
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and <t>β-tubulin</t> were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Pstat3 S727 Pstat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mx1  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc mx1
SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and <t>β-tubulin</t> were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.
Mx1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Journal: Journal of Interferon & Cytokine Research

Article Title: Type I Interferon-Mediated Induction of Antiviral Genes and Proteins Fails to Protect Cells from the Cytopathic Effects of Sendai Virus Infection

doi: 10.1089/jir.2016.0051

Figure Lengend Snippet: SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Article Snippet: Antibodies and dilutions The following antibodies and dilutions were used for the western blot analyses: Monoclonal rabbit anti-pIRF-3(S386; Abcam, Cambridge, MA): 1:5,000; monoclonal anti-IRF-3 (Cell Signaling Technologies, Danvers, MA): 1:1,000; polyclonal rabbit anti-IRF-7 (Santa Cruz Biotechnologies, Dallas, TX): 1:1,000; polyclonal rabbit anti-STAT1 (Santa Cruz Biotechnologies): 1:1,000; monoclonal mouse anti-pSTAT1(Y701; BD Transduction Laboratories, San Jose, CA): 1:5,000; polyclonal rabbit anti-STAT2 (Santa Cruz Biotechnologies): 1:1,000; polyclonal rabbit anti-pSTAT2(Y689; End Millipore, Billerica, MA): 1:2,000; monoclonal mouse anti-IRF-9(ISGF3γ; BD Transduction Laboratories): 1:1,000; monoclonal mouse anti-Actin (Cell Signaling Technologies): 1:5,000; polyclonal rabbit anti-β tubulin (Cell Signaling Technologies): 1:2,000; mouse monoclonal RNA polymerase II (Santa Cruz Biotechnologies): 1:1,000; rabbit polyclonal IFIT3 (Covance, Inc., Denver, PA) 1:1,000; rabbit polyclonal ISG15: 1:1,000 (Santa Cruz Biotechnologies); mouse polyclonal Mx1: 1:1,000 (Abnova, Taipei, Taiwan).

Techniques: Infection, Western Blot, Expressing

SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Journal: Journal of Interferon & Cytokine Research

Article Title: Type I Interferon-Mediated Induction of Antiviral Genes and Proteins Fails to Protect Cells from the Cytopathic Effects of Sendai Virus Infection

doi: 10.1089/jir.2016.0051

Figure Lengend Snippet: SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Article Snippet: The following antibodies and dilutions were used for the western blot analyses: Monoclonal rabbit anti-pIRF-3(S386; Abcam, Cambridge, MA): 1:5,000; monoclonal anti-IRF-3 (Cell Signaling Technologies, Danvers, MA): 1:1,000; polyclonal rabbit anti-IRF-7 (Santa Cruz Biotechnologies, Dallas, TX): 1:1,000; polyclonal rabbit anti-STAT1 (Santa Cruz Biotechnologies): 1:1,000; monoclonal mouse anti-pSTAT1(Y701; BD Transduction Laboratories, San Jose, CA): 1:5,000; polyclonal rabbit anti-STAT2 (Santa Cruz Biotechnologies): 1:1,000; polyclonal rabbit anti-pSTAT2(Y689; End Millipore, Billerica, MA): 1:2,000; monoclonal mouse anti-IRF-9(ISGF3γ; BD Transduction Laboratories): 1:1,000; monoclonal mouse anti-Actin (Cell Signaling Technologies): 1:5,000; polyclonal rabbit anti-β tubulin (Cell Signaling Technologies): 1:2,000; mouse monoclonal RNA polymerase II (Santa Cruz Biotechnologies): 1:1,000; rabbit polyclonal IFIT3 (Covance, Inc., Denver, PA) 1:1,000; rabbit polyclonal ISG15: 1:1,000 (Santa Cruz Biotechnologies); mouse polyclonal Mx1: 1:1,000 (Abnova, Taipei, Taiwan).

Techniques: Infection, Western Blot, Expressing

Inhibiting canonical type I IFN signaling during SeV infection does not affect SeV-mediated CPE. (A) U937 cells were treated with 1000 U/mL of IFN-α2a, IFN-β, or IFN-ω for 1 h in the presence or absence of the IFNAR2 neutralizing antibody. Western blot analysis was performed to detect phosphorylated STAT1(Y701) and tubulin was used as the loading control. UT: untreated; NA: cells treated with IFNAR2 neutralizing antibody alone; α2a: IFN-α2a treated; α2a + NA: cells treated with both antibody and IFN-α2a; β: IFN-β treated; β + NA: cells treated with both antibody and IFN-β; ω: IFN-ω treated; and ω + NA: cells treated with both antibody and IFN-ω. (B) U937 (panel 1) and A549 cells (panel 2) were infected with SeV (150 HA U/mL) in the presence or absence of IFNAR2 neutralizing antibody. Cell viability was measured 72 h pi. Data are representative of 3 independent biological replicates. Supernatants from cells infected with SeV (150 HA U/mL) for 16 h were inactivated (β-propiolactone) and used to pretreat fresh (C) U937 and (D) A549 cells for 24 h in the presence or absence of the IFNAR or IFNLR neutralizing antibodies. The U937 cells were then infected with either EMCV or SeV (150 HA U/mL) for 72 h and cell viability was measured via MTT assay (U937) or crystal violet assay (A549). Results are shown as the % of cell control. SeV150: virus-inactivated supernatant from SeV-infected cells (150 HA U/mL); SeV150 + IFNAR2 NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNAR2-neutralizing antibody; SeV150 + IFNLR NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNLR-neutralizing antibody. IFNAR, IFN receptor; IFNLR, IFN-lambda receptor.

Journal: Journal of Interferon & Cytokine Research

Article Title: Type I Interferon-Mediated Induction of Antiviral Genes and Proteins Fails to Protect Cells from the Cytopathic Effects of Sendai Virus Infection

doi: 10.1089/jir.2016.0051

Figure Lengend Snippet: Inhibiting canonical type I IFN signaling during SeV infection does not affect SeV-mediated CPE. (A) U937 cells were treated with 1000 U/mL of IFN-α2a, IFN-β, or IFN-ω for 1 h in the presence or absence of the IFNAR2 neutralizing antibody. Western blot analysis was performed to detect phosphorylated STAT1(Y701) and tubulin was used as the loading control. UT: untreated; NA: cells treated with IFNAR2 neutralizing antibody alone; α2a: IFN-α2a treated; α2a + NA: cells treated with both antibody and IFN-α2a; β: IFN-β treated; β + NA: cells treated with both antibody and IFN-β; ω: IFN-ω treated; and ω + NA: cells treated with both antibody and IFN-ω. (B) U937 (panel 1) and A549 cells (panel 2) were infected with SeV (150 HA U/mL) in the presence or absence of IFNAR2 neutralizing antibody. Cell viability was measured 72 h pi. Data are representative of 3 independent biological replicates. Supernatants from cells infected with SeV (150 HA U/mL) for 16 h were inactivated (β-propiolactone) and used to pretreat fresh (C) U937 and (D) A549 cells for 24 h in the presence or absence of the IFNAR or IFNLR neutralizing antibodies. The U937 cells were then infected with either EMCV or SeV (150 HA U/mL) for 72 h and cell viability was measured via MTT assay (U937) or crystal violet assay (A549). Results are shown as the % of cell control. SeV150: virus-inactivated supernatant from SeV-infected cells (150 HA U/mL); SeV150 + IFNAR2 NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNAR2-neutralizing antibody; SeV150 + IFNLR NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNLR-neutralizing antibody. IFNAR, IFN receptor; IFNLR, IFN-lambda receptor.

Article Snippet: The following antibodies and dilutions were used for the western blot analyses: Monoclonal rabbit anti-pIRF-3(S386; Abcam, Cambridge, MA): 1:5,000; monoclonal anti-IRF-3 (Cell Signaling Technologies, Danvers, MA): 1:1,000; polyclonal rabbit anti-IRF-7 (Santa Cruz Biotechnologies, Dallas, TX): 1:1,000; polyclonal rabbit anti-STAT1 (Santa Cruz Biotechnologies): 1:1,000; monoclonal mouse anti-pSTAT1(Y701; BD Transduction Laboratories, San Jose, CA): 1:5,000; polyclonal rabbit anti-STAT2 (Santa Cruz Biotechnologies): 1:1,000; polyclonal rabbit anti-pSTAT2(Y689; End Millipore, Billerica, MA): 1:2,000; monoclonal mouse anti-IRF-9(ISGF3γ; BD Transduction Laboratories): 1:1,000; monoclonal mouse anti-Actin (Cell Signaling Technologies): 1:5,000; polyclonal rabbit anti-β tubulin (Cell Signaling Technologies): 1:2,000; mouse monoclonal RNA polymerase II (Santa Cruz Biotechnologies): 1:1,000; rabbit polyclonal IFIT3 (Covance, Inc., Denver, PA) 1:1,000; rabbit polyclonal ISG15: 1:1,000 (Santa Cruz Biotechnologies); mouse polyclonal Mx1: 1:1,000 (Abnova, Taipei, Taiwan).

Techniques: Infection, Western Blot, MTT Assay, Crystal Violet Assay

SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Journal: Journal of Interferon & Cytokine Research

Article Title: Type I Interferon-Mediated Induction of Antiviral Genes and Proteins Fails to Protect Cells from the Cytopathic Effects of Sendai Virus Infection

doi: 10.1089/jir.2016.0051

Figure Lengend Snippet: SeV-induced IFN production activates Jak/STAT signaling and ISG induction. (A) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. Nuclear and cytoplasmic protein extracts were prepared and western blots probing for pIRF-3(S386), IRF-3, IRF-7, and IRF-9 were performed. RNA pol II and β-tubulin were the nuclear and cytoplasmic loading controls, respectively. N, nuclear, C, cytoplasmic. (B) U937 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole cell protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), ISG15, and IFIT3, and tubulin as the loading control were performed. (C) A549 cells were infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h and whole protein lysates were prepared. Western blots probing for pSTAT1(Y701), STAT1(total), pSTAT2(Y689), STAT2(total), IRF-9, IRF-7, and Mx1, and tubulin as the loading control were performed. (D) ISG15 and IFIT3 mRNA expression in U937 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. (E) IRF-7 and Mx1 mRNA expression in A549 cells infected with SeV (150 HA U/mL) for 4, 8, 16, and 24 h. IRF-7 and Mx1 mRNA induction in (F) primary monocytes and (G) Namalwa cells infected with SeV (150 HA U/mL) for 24 h. All gene induction data are expressed as the % of relative HPRT expression. All data are representative of 3 independent biological replicates. UI, uninfected, untreated. IRF, IFN regulatory factor; ISG, interferon-stimulated gene; HPRT, hypoxanthine-guanine phosphoribosyltransferase.

Article Snippet: The following antibodies and dilutions were used for the western blot analyses: Monoclonal rabbit anti-pIRF-3(S386; Abcam, Cambridge, MA): 1:5,000; monoclonal anti-IRF-3 (Cell Signaling Technologies, Danvers, MA): 1:1,000; polyclonal rabbit anti-IRF-7 (Santa Cruz Biotechnologies, Dallas, TX): 1:1,000; polyclonal rabbit anti-STAT1 (Santa Cruz Biotechnologies): 1:1,000; monoclonal mouse anti-pSTAT1(Y701; BD Transduction Laboratories, San Jose, CA): 1:5,000; polyclonal rabbit anti-STAT2 (Santa Cruz Biotechnologies): 1:1,000; polyclonal rabbit anti-pSTAT2(Y689; End Millipore, Billerica, MA): 1:2,000; monoclonal mouse anti-IRF-9(ISGF3γ; BD Transduction Laboratories): 1:1,000; monoclonal mouse anti-Actin (Cell Signaling Technologies): 1:5,000; polyclonal rabbit anti-β tubulin (Cell Signaling Technologies): 1:2,000; mouse monoclonal RNA polymerase II (Santa Cruz Biotechnologies): 1:1,000; rabbit polyclonal IFIT3 (Covance, Inc., Denver, PA) 1:1,000; rabbit polyclonal ISG15: 1:1,000 (Santa Cruz Biotechnologies); mouse polyclonal Mx1: 1:1,000 (Abnova, Taipei, Taiwan).

Techniques: Infection, Western Blot, Control, Expressing

Inhibiting canonical type I IFN signaling during SeV infection does not affect SeV-mediated CPE. (A) U937 cells were treated with 1000 U/mL of IFN-α2a, IFN-β, or IFN-ω for 1 h in the presence or absence of the IFNAR2 neutralizing antibody. Western blot analysis was performed to detect phosphorylated STAT1(Y701) and tubulin was used as the loading control. UT: untreated; NA: cells treated with IFNAR2 neutralizing antibody alone; α2a: IFN-α2a treated; α2a + NA: cells treated with both antibody and IFN-α2a; β: IFN-β treated; β + NA: cells treated with both antibody and IFN-β; ω: IFN-ω treated; and ω + NA: cells treated with both antibody and IFN-ω. (B) U937 (panel 1) and A549 cells (panel 2) were infected with SeV (150 HA U/mL) in the presence or absence of IFNAR2 neutralizing antibody. Cell viability was measured 72 h pi. Data are representative of 3 independent biological replicates. Supernatants from cells infected with SeV (150 HA U/mL) for 16 h were inactivated (β-propiolactone) and used to pretreat fresh (C) U937 and (D) A549 cells for 24 h in the presence or absence of the IFNAR or IFNLR neutralizing antibodies. The U937 cells were then infected with either EMCV or SeV (150 HA U/mL) for 72 h and cell viability was measured via MTT assay (U937) or crystal violet assay (A549). Results are shown as the % of cell control. SeV150: virus-inactivated supernatant from SeV-infected cells (150 HA U/mL); SeV150 + IFNAR2 NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNAR2-neutralizing antibody; SeV150 + IFNLR NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNLR-neutralizing antibody. IFNAR, IFN receptor; IFNLR, IFN-lambda receptor.

Journal: Journal of Interferon & Cytokine Research

Article Title: Type I Interferon-Mediated Induction of Antiviral Genes and Proteins Fails to Protect Cells from the Cytopathic Effects of Sendai Virus Infection

doi: 10.1089/jir.2016.0051

Figure Lengend Snippet: Inhibiting canonical type I IFN signaling during SeV infection does not affect SeV-mediated CPE. (A) U937 cells were treated with 1000 U/mL of IFN-α2a, IFN-β, or IFN-ω for 1 h in the presence or absence of the IFNAR2 neutralizing antibody. Western blot analysis was performed to detect phosphorylated STAT1(Y701) and tubulin was used as the loading control. UT: untreated; NA: cells treated with IFNAR2 neutralizing antibody alone; α2a: IFN-α2a treated; α2a + NA: cells treated with both antibody and IFN-α2a; β: IFN-β treated; β + NA: cells treated with both antibody and IFN-β; ω: IFN-ω treated; and ω + NA: cells treated with both antibody and IFN-ω. (B) U937 (panel 1) and A549 cells (panel 2) were infected with SeV (150 HA U/mL) in the presence or absence of IFNAR2 neutralizing antibody. Cell viability was measured 72 h pi. Data are representative of 3 independent biological replicates. Supernatants from cells infected with SeV (150 HA U/mL) for 16 h were inactivated (β-propiolactone) and used to pretreat fresh (C) U937 and (D) A549 cells for 24 h in the presence or absence of the IFNAR or IFNLR neutralizing antibodies. The U937 cells were then infected with either EMCV or SeV (150 HA U/mL) for 72 h and cell viability was measured via MTT assay (U937) or crystal violet assay (A549). Results are shown as the % of cell control. SeV150: virus-inactivated supernatant from SeV-infected cells (150 HA U/mL); SeV150 + IFNAR2 NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNAR2-neutralizing antibody; SeV150 + IFNLR NA: treated with virus-inactivated supernatant from SeV-infected cells (150 HA U/mL) in the presence of IFNLR-neutralizing antibody. IFNAR, IFN receptor; IFNLR, IFN-lambda receptor.

Article Snippet: The following antibodies and dilutions were used for the western blot analyses: Monoclonal rabbit anti-pIRF-3(S386; Abcam, Cambridge, MA): 1:5,000; monoclonal anti-IRF-3 (Cell Signaling Technologies, Danvers, MA): 1:1,000; polyclonal rabbit anti-IRF-7 (Santa Cruz Biotechnologies, Dallas, TX): 1:1,000; polyclonal rabbit anti-STAT1 (Santa Cruz Biotechnologies): 1:1,000; monoclonal mouse anti-pSTAT1(Y701; BD Transduction Laboratories, San Jose, CA): 1:5,000; polyclonal rabbit anti-STAT2 (Santa Cruz Biotechnologies): 1:1,000; polyclonal rabbit anti-pSTAT2(Y689; End Millipore, Billerica, MA): 1:2,000; monoclonal mouse anti-IRF-9(ISGF3γ; BD Transduction Laboratories): 1:1,000; monoclonal mouse anti-Actin (Cell Signaling Technologies): 1:5,000; polyclonal rabbit anti-β tubulin (Cell Signaling Technologies): 1:2,000; mouse monoclonal RNA polymerase II (Santa Cruz Biotechnologies): 1:1,000; rabbit polyclonal IFIT3 (Covance, Inc., Denver, PA) 1:1,000; rabbit polyclonal ISG15: 1:1,000 (Santa Cruz Biotechnologies); mouse polyclonal Mx1: 1:1,000 (Abnova, Taipei, Taiwan).

Techniques: Infection, Western Blot, Control, MTT Assay, Crystal Violet Assay, Virus