rabbit anti ifnar2 pab Search Results


rea124 miltenyi biotec  (Miltenyi Biotec)
94
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anti ifnar2 antibodies  (Sino Biological)
92
Sino Biological anti ifnar2 antibodies
Primer sequences for quantitative real-time PCR.
Anti Ifnar2 Antibodies, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti ifnar2 pab  (Proteintech)
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Proteintech rabbit anti ifnar2 pab
Primer sequences for quantitative real-time PCR.
Rabbit Anti Ifnar2 Pab, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti-human ifnar2  (PBL Biomedical Laboratories)
90
PBL Biomedical Laboratories rabbit anti-human ifnar2
Primer sequences for quantitative real-time PCR.
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anti ifnar2 fitc  (Sino Biological)
90
Sino Biological anti ifnar2 fitc
(a) U6A cells, stably transduced with control (−) or C-terminally Myc-tagged STAT2 (STAT2-Myc), were infected with MIP control (−) or MIP-USP18 (+) retrovirus. With or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (b–d) U6A cells, stably transduced to express either full-length STAT2 (STAT2-Myc), or the indicated STAT2 deletion mutant (b, STAT2ΔCC-Myc; c, STAT2ΔDB-Myc; d, STAT2ΔCC/DB-Myc) with or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (e) Histograms showing the surface expression of IFNAR1 and <t>IFNAR2</t> following infection with indicated constructs in U6A cell lines.
Anti Ifnar2 Fitc, supplied by Sino Biological, 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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anti human ifnar2  (Miltenyi Biotec)
94
Miltenyi Biotec anti human ifnar2
Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and <t>IFNAR2</t> gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.
Anti Human Ifnar2, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti kshv orf 45  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc mouse anti kshv orf 45
Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and <t>IFNAR2</t> gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.
Mouse Anti Kshv Orf 45, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal igg sc-704 against human ifnar2 antibody  (Santa Cruz Biotechnology)
90
Santa Cruz Biotechnology rabbit polyclonal igg sc-704 against human ifnar2 antibody
Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and <t>IFNAR2</t> gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.
Rabbit Polyclonal Igg Sc 704 Against Human Ifnar2 Antibody, supplied by Santa Cruz Biotechnology, 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 anti-ifnar2  (Proteintech)
90
Proteintech rabbit anti-ifnar2
Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and <t>IFNAR2</t> gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.
Rabbit Anti Ifnar2, supplied by Proteintech, 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 anti glutathione peroxidase 4  (Boster Bio)
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Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and <t>IFNAR2</t> gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.
Rabbit Anti Glutathione Peroxidase 4, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal anti-human ifnar2 antibody  (Abnova)
90
Abnova rabbit polyclonal anti-human ifnar2 antibody
Longitudinal assessment of IFNAR1, <t>IFNAR2,</t> sIFNAR2 and MxA gene expression. RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from MS patients before and after 6 months of IFN-β therapy onset (N=41), assessed by real time-PCR using GAPDH as raeference gene. Significant p-values are shown with asterisk.
Rabbit Polyclonal Anti Human Ifnar2 Antibody, 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 anti interferon alpha beta receptor 1 pab  (Danaher Inc)
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Danaher Inc rabbit anti interferon alpha beta receptor 1 pab
Longitudinal assessment of IFNAR1, <t>IFNAR2,</t> sIFNAR2 and MxA gene expression. RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from MS patients before and after 6 months of IFN-β therapy onset (N=41), assessed by real time-PCR using GAPDH as raeference gene. Significant p-values are shown with asterisk.
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Image Search Results


Primer sequences for quantitative real-time PCR.

Journal: Molecular Medicine Reports

Article Title: Influenza virus non-structural protein 1 inhibits the production of interferon β of alveolar epithelial cells upon the infection of influenza A H1N1

doi: 10.3892/mmr.2017.7138

Figure Lengend Snippet: Primer sequences for quantitative real-time PCR.

Article Snippet: FITC-conjugated anti-IFNAR2 antibodies and isotype antibodies were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Sequencing

NS1 downregulates the expression of receptors for IFNβ-IFNAR1 and IFNAR2. (A and B) The expression of Ifnar1 and Ifnar2 was assessed by real-time q-PCR. NS1-expressing A549 cells produced lower amounts of mRNAs of IFNAR1 and IFNAR2 (P<0.001, n=4). (C) The protein levels of IFNAR1 and IFNAR2 were measured by western blot analysis. NS1 inhibited the upregulation of IFNAR1 and IFNAR2 of A549 cells upon H1N1 infection. (D) The surface expression of IFNAR1 and IFNAR2 on infected A549 cells were checked by flow cytometry. NS1-expressing A549 cells (blue curves) had decreased levels of IFN receptors in A549 cells infected with H1N1. (E) The proportion of IFNAR1-expressing cells of infected A549 cells with or without NS1 expression measured by flow cytometry, P<0.001, n=4. (F) The percentage of IFNAR2-expressing cells infected with H1N1, P<0.001, n=4. *P<0.05; ***P<0.001. NS1, non-structural protein 1; IFN, interferon; HA, haemagglutination tests; n.s., non significant.

Journal: Molecular Medicine Reports

Article Title: Influenza virus non-structural protein 1 inhibits the production of interferon β of alveolar epithelial cells upon the infection of influenza A H1N1

doi: 10.3892/mmr.2017.7138

Figure Lengend Snippet: NS1 downregulates the expression of receptors for IFNβ-IFNAR1 and IFNAR2. (A and B) The expression of Ifnar1 and Ifnar2 was assessed by real-time q-PCR. NS1-expressing A549 cells produced lower amounts of mRNAs of IFNAR1 and IFNAR2 (P<0.001, n=4). (C) The protein levels of IFNAR1 and IFNAR2 were measured by western blot analysis. NS1 inhibited the upregulation of IFNAR1 and IFNAR2 of A549 cells upon H1N1 infection. (D) The surface expression of IFNAR1 and IFNAR2 on infected A549 cells were checked by flow cytometry. NS1-expressing A549 cells (blue curves) had decreased levels of IFN receptors in A549 cells infected with H1N1. (E) The proportion of IFNAR1-expressing cells of infected A549 cells with or without NS1 expression measured by flow cytometry, P<0.001, n=4. (F) The percentage of IFNAR2-expressing cells infected with H1N1, P<0.001, n=4. *P<0.05; ***P<0.001. NS1, non-structural protein 1; IFN, interferon; HA, haemagglutination tests; n.s., non significant.

Article Snippet: FITC-conjugated anti-IFNAR2 antibodies and isotype antibodies were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Expressing, Produced, Western Blot, Infection, Flow Cytometry

(a) U6A cells, stably transduced with control (−) or C-terminally Myc-tagged STAT2 (STAT2-Myc), were infected with MIP control (−) or MIP-USP18 (+) retrovirus. With or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (b–d) U6A cells, stably transduced to express either full-length STAT2 (STAT2-Myc), or the indicated STAT2 deletion mutant (b, STAT2ΔCC-Myc; c, STAT2ΔDB-Myc; d, STAT2ΔCC/DB-Myc) with or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (e) Histograms showing the surface expression of IFNAR1 and IFNAR2 following infection with indicated constructs in U6A cell lines.

Journal: Nature structural & molecular biology

Article Title: STAT2 is an essential adaptor in USP18-mediated suppression of type I interferon signaling

doi: 10.1038/nsmb.3378

Figure Lengend Snippet: (a) U6A cells, stably transduced with control (−) or C-terminally Myc-tagged STAT2 (STAT2-Myc), were infected with MIP control (−) or MIP-USP18 (+) retrovirus. With or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (b–d) U6A cells, stably transduced to express either full-length STAT2 (STAT2-Myc), or the indicated STAT2 deletion mutant (b, STAT2ΔCC-Myc; c, STAT2ΔDB-Myc; d, STAT2ΔCC/DB-Myc) with or without IFNα (1000 U/ml) treatment for 15 minutes cell lysates were collected and analyzed by Western blotting with the indicated antibodies. (e) Histograms showing the surface expression of IFNAR1 and IFNAR2 following infection with indicated constructs in U6A cell lines.

Article Snippet: Anti-IFNAR2 FITC was purchased from Sino Biological Inc. Anti-USP18 antibody is described as before .

Techniques: Stable Transfection, Transduction, Infection, Western Blot, Mutagenesis, Expressing, Construct

(a) IB analysis of WCL and anti-FLAG IP derived from U6A cells 24 hours after co-transfection with plasmids encoding USP18, FLAG-IFNAR2, and increasing amounts of STAT2-Myc expression construct. The relative USP18 binding to IFNAR2 from three independent experiments was quantified and plotted as the ratio of IFNAR2-bound USP18 to total USP18 (right panel). Data are normalized to the maximum binding (lane 4). (b) Recruitment of USP18 and STAT2 to micropatterned IFNAR2 in STAT2-deficient U6A cells as illustrated in the cartoon. U6A cells transfected with HaloTag-mTagBFP-IFNAR2 and mEGFP-USP18 (USP18 –STAT2) (green channel, left image) and U6A cells transfected with HaloTag-mTagBFP-IFNAR2, mEGFP-USP18 and STAT2-TagRFP-T (USP18 +STAT2) (green channel, right image). Representative images of 17 cells analyzed in two independent experiments. (c) Recruitment of USP18 to immobilized IFNAR2 (R2) into micropatterns was quantified in U6A cells by determining the contrast of the fluorescence intensities inside and outside the patterns. For comparison, constitutive binding of STAT2 (positive control) and cytosolic mEGFP (negative control) to micropatterned full length IFNAR2 expressed in U6A cells was quantified. n: number of cells analyzed in two independent experiments. Significance was quantified using the two-sample Kolmogorov-Smirnov test. *** P < 0.001, n.s., not significant. (d) The C-terminal STAT2 interacting region of IFNAR2 is necessary for recruiting USP18. Quantification of recruitment of mEGFP-USP18 co-expressed with STAT2-TagRFP-T to immobilized HaloTag-IFNAR2 or C-terminally truncated HaloTag-IFNAR2 (R2Δ375) in Hela cells. As negative controls, HeLa cells were transfected with cytosolic mEGFP or TagRFP-T, respectively. n: number of cells analyzed in two independent experiments. Significance was quantified using the two-sample Kolmogorov-Smirnov test. *** P < 0.001, n.s. not significant.

Journal: Nature structural & molecular biology

Article Title: STAT2 is an essential adaptor in USP18-mediated suppression of type I interferon signaling

doi: 10.1038/nsmb.3378

Figure Lengend Snippet: (a) IB analysis of WCL and anti-FLAG IP derived from U6A cells 24 hours after co-transfection with plasmids encoding USP18, FLAG-IFNAR2, and increasing amounts of STAT2-Myc expression construct. The relative USP18 binding to IFNAR2 from three independent experiments was quantified and plotted as the ratio of IFNAR2-bound USP18 to total USP18 (right panel). Data are normalized to the maximum binding (lane 4). (b) Recruitment of USP18 and STAT2 to micropatterned IFNAR2 in STAT2-deficient U6A cells as illustrated in the cartoon. U6A cells transfected with HaloTag-mTagBFP-IFNAR2 and mEGFP-USP18 (USP18 –STAT2) (green channel, left image) and U6A cells transfected with HaloTag-mTagBFP-IFNAR2, mEGFP-USP18 and STAT2-TagRFP-T (USP18 +STAT2) (green channel, right image). Representative images of 17 cells analyzed in two independent experiments. (c) Recruitment of USP18 to immobilized IFNAR2 (R2) into micropatterns was quantified in U6A cells by determining the contrast of the fluorescence intensities inside and outside the patterns. For comparison, constitutive binding of STAT2 (positive control) and cytosolic mEGFP (negative control) to micropatterned full length IFNAR2 expressed in U6A cells was quantified. n: number of cells analyzed in two independent experiments. Significance was quantified using the two-sample Kolmogorov-Smirnov test. *** P < 0.001, n.s., not significant. (d) The C-terminal STAT2 interacting region of IFNAR2 is necessary for recruiting USP18. Quantification of recruitment of mEGFP-USP18 co-expressed with STAT2-TagRFP-T to immobilized HaloTag-IFNAR2 or C-terminally truncated HaloTag-IFNAR2 (R2Δ375) in Hela cells. As negative controls, HeLa cells were transfected with cytosolic mEGFP or TagRFP-T, respectively. n: number of cells analyzed in two independent experiments. Significance was quantified using the two-sample Kolmogorov-Smirnov test. *** P < 0.001, n.s. not significant.

Article Snippet: Anti-IFNAR2 FITC was purchased from Sino Biological Inc. Anti-USP18 antibody is described as before .

Techniques: Derivative Assay, Cotransfection, Expressing, Construct, Binding Assay, Transfection, Fluorescence, Positive Control, Negative Control

Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and IFNAR2 gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.

Journal: Nature Communications

Article Title: Type I interferon signaling induces melanoma cell-intrinsic PD-1 and its inhibition antagonizes immune checkpoint blockade

doi: 10.1038/s41467-024-51496-2

Figure Lengend Snippet: Single-cell RNA-seq analysis of a type I interferon-α/β receptor (IFNAR) and type II interferon-γ receptor (IFNGR) subunit gene expression shown in units of transcript per million (TPM)/10 (black bars; mean ± SEM) in patient melanoma cells (left) versus tumor-infiltrating T-cells (right). Gene expression of additional cytokine and growth factor receptor subunits is also shown (gray bars). b Violin plots of IFNAR1 and IFNAR2 gene expression (TPM/10; median, bold white line; top and bottom quartiles, thin white lines) in patient melanoma (MM) versus tumor-infiltrating T-cells generated from the single-cell RNA-seq dataset above. c IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by patient tumor cells and tumor-infiltrating lymphocytes (TILs), with representative flow cytometric histograms shown (right). d Relative IFNAR1 and IFNAR2 gene expression (fold change, mean ± SEM) in human melanoma cell lines (black bars) versus human activated CD3 + T-cells (gray bars) as determined by RT-qPCR. e IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by human A2058 and A375 melanoma cells, with representative flow cytometric histograms shown for A2058 melanoma cells (right). f Relative Ifnar1 and Ifnar2 gene expression (fold change, mean ± SEM) in murine melanoma cell lines (black bars) versus murine activated CD3 + T-cells (gray bars) as determined by RT-qPCR. g IFNAR1 and IFNAR2 surface protein expression (percent positivity, mean ± SEM, left) by murine melanoma cells, with representative flow cytometric histograms shown for B16-F10 and YUMM1.7 melanoma cells (right). IFNAR1 (left) and IFNAR2 (right) surface protein expression (percent positivity, mean ± SEM) by PD-1 + versus PD-1 - h , human A2058 and A375 and i , murine melanoma cell subsets, with representative flow cytometric histograms shown for A2058 cells. Results represent biologically independent samples of a , b n = 1252 MM cells (left panel) and 2040 T-cells (right panel), c n = 4, and biologically independent experiments of d , f , i n = 3, e n = 12, g n = 6, and h n = 7 (A2058) and n = 6 (A375). Statistical analyses included b Mann-Whitney test, two-sided, c paired t -test, one-sided, and h , i unpaired t -test, two-sided. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, not significant. See also Supplementary Figs. – . Source data, including exact p -values, are provided as a Source Data file.

Article Snippet: The following antibodies (abs) were used for flow cytometric analyses of human cells: Alexa Fluor 647-conjugated (BD Biosciences) or PerCP-eFluor 710-conjugated anti-human PD-1 (clone MIH4, 20 μg/mL, Thermo Fisher Scientific, Waltham, MA) and Alexa Fluor 647-conjugated or PerCP-eFluor 710-conjugated mouse IgG1 isotype controls (clone MOPC-31C, 20 μg/mL, BD Biosciences; clone P3.6.2.8.1, 20 μg/mL, Thermo Fisher Scientific), unconjugated anti-human PD-1 clinical ab, nivolumab (100 μg/mL, Bristol Myers Squibb, Cambridge, MA) was obtained from the BWH Pharmacy, and Ultra-LEAF unconjugated human IgG4 isotype control (clone QA16A15, 100 μg/mL, BioLegend), FITC-conjugated or PE-conjugated anti-human IgG4 (clone HP6023, 1:50, Abcam, Waltham, MA), PE-conjugated anti-human PD-L1 (clone 29E.2A3, 15 μg/mL, BioLegend) and PE-conjugated mouse IgG2b isotype control (clone MPC-11, 15 μg/mL, BioLegend), APC-conjugated anti-human PD-L2 (clone 24 F.10C12, 15 μg/mL, BioLegend) and APC-conjugated mouse IgG2a isotype control (clone MOPC-173, 15 μg/mL, BioLegend), PE-conjugated anti-human IFNAR1 (clone 85228, 20 μg/mL, Thermo Fisher Scientific) and PE-conjugated mouse IgG1 isotype control (clone P3.6.2.8.1, 20 μg/mL, Thermo Fisher Scientific) or PE-conjugated mouse IgG1 isotype control (clone MOPC-21, 20 μg/mL, BioLegend), Alexa Fluor 750-conjugated anti-human IFNAR1 (clone 85228, 20 μg/mL, R&D Systems, Minneapolis, MN) and Alexa Fluor 750-conjugated mouse IgG1 isotype control (clone 11711, 20 μg/mL, R&D Systems), APC-conjugated or APC-Vio770-conjugated anti-human IFNAR2 (clone REA124, 20 μg/mL, Miltenyi Biotec, Gaithersburg, MD) and APC-conjugated or APC-Vio770-conjugated human IgG1 isotype control (clone REA293, 20 μg/mL, Miltenyi Biotec), FITC-conjugated anti-human IFN-β (clone A1(IFNb)), 5 μg/mL, and FITC-conjugated mouse IgG1 (clone MOPC-21, 5 μg/mL, Thermo Fisher Scientific), PE-conjugated anti-human IFN-α[2b] (clone 7N4-1, 1.5 μg/mL) and PE-conjugated mouse IgG1 isotype control (clone MOPC-21, 1.5 μg/mL, BD Biosciences), FITC-conjugated anti-mouse Interferon alpha (clone RMMA-1, 50 μg/mL, PBL Assay Science, Piscataway, NJ) and FITC-conjugated rat IgG1 (clone eBRG1, 50 μg/mL, Thermo Fisher Scientific), anti-mouse Interferon-beta 1 (clone D2J1D, 75 ng/mL), rabbit IgG isotype control (clone DA1E, 75 ng/mL Cell Signaling Technology, Danvers, MA), and PE-conjugated donkey anti-rabbit IgG (clone Poly4064, 1.25 μg/mL BioLegend).

Techniques: RNA Sequencing, Gene Expression, Generated, Expressing, Quantitative RT-PCR, MANN-WHITNEY

Longitudinal assessment of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression. RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from MS patients before and after 6 months of IFN-β therapy onset (N=41), assessed by real time-PCR using GAPDH as raeference gene. Significant p-values are shown with asterisk.

Journal: Frontiers in Immunology

Article Title: Soluble Receptor Isoform of IFN-Beta (sIFNAR2) in Multiple Sclerosis Patients and Their Association With the Clinical Response to IFN-Beta Treatment

doi: 10.3389/fimmu.2021.778204

Figure Lengend Snippet: Longitudinal assessment of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression. RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from MS patients before and after 6 months of IFN-β therapy onset (N=41), assessed by real time-PCR using GAPDH as raeference gene. Significant p-values are shown with asterisk.

Article Snippet: Briefly, the plates were coated with 0.2-μg rabbit polyclonal anti-human IFNAR2 antibody (Abnova) overnight.

Techniques: Gene Expression, Expressing, Real-time Polymerase Chain Reaction

IFNAR2 splice variants under IFN-β treatment. (A) Correlation analysis between the transmembrane (IFNAR2) and soluble (sIFNAR2) transcripts before (r=0.636 p< 0.001) and after 6 months of IFN-β therapy onset (r=0.749 p<0.001). (B) Comparison of the relative expression of IFNAR2 versus sIFNAR2 before and after 6 months of IFN-β therapy onset, using GAPDH as a raeference gene (N=41). Significant p-values are shown with asterisk.

Journal: Frontiers in Immunology

Article Title: Soluble Receptor Isoform of IFN-Beta (sIFNAR2) in Multiple Sclerosis Patients and Their Association With the Clinical Response to IFN-Beta Treatment

doi: 10.3389/fimmu.2021.778204

Figure Lengend Snippet: IFNAR2 splice variants under IFN-β treatment. (A) Correlation analysis between the transmembrane (IFNAR2) and soluble (sIFNAR2) transcripts before (r=0.636 p< 0.001) and after 6 months of IFN-β therapy onset (r=0.749 p<0.001). (B) Comparison of the relative expression of IFNAR2 versus sIFNAR2 before and after 6 months of IFN-β therapy onset, using GAPDH as a raeference gene (N=41). Significant p-values are shown with asterisk.

Article Snippet: Briefly, the plates were coated with 0.2-μg rabbit polyclonal anti-human IFNAR2 antibody (Abnova) overnight.

Techniques: Comparison, Expressing

Longitudinal assessment of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression in responder and non-responder patients. (A) Heat map representing the gene expression data of IFNAR1, IFNAR2, sIFNAR2, and MxA between responder and non-responder patients, at baseline and 6 months after the onset of IFN-β therapy. Unchanged proteins are displayed in black, up-regulated proteins are displayed in green while the down-regulated proteins are depicted in red. (B) Bar chart representing the gene expression for responders (N=22) and non-responders (N=19) using GAPDH as raeference gene. Significant p-values are shown with asterisk.

Journal: Frontiers in Immunology

Article Title: Soluble Receptor Isoform of IFN-Beta (sIFNAR2) in Multiple Sclerosis Patients and Their Association With the Clinical Response to IFN-Beta Treatment

doi: 10.3389/fimmu.2021.778204

Figure Lengend Snippet: Longitudinal assessment of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression in responder and non-responder patients. (A) Heat map representing the gene expression data of IFNAR1, IFNAR2, sIFNAR2, and MxA between responder and non-responder patients, at baseline and 6 months after the onset of IFN-β therapy. Unchanged proteins are displayed in black, up-regulated proteins are displayed in green while the down-regulated proteins are depicted in red. (B) Bar chart representing the gene expression for responders (N=22) and non-responders (N=19) using GAPDH as raeference gene. Significant p-values are shown with asterisk.

Article Snippet: Briefly, the plates were coated with 0.2-μg rabbit polyclonal anti-human IFNAR2 antibody (Abnova) overnight.

Techniques: Gene Expression

In vitro study of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression. Representation of RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from untreated MS patients cultured with or without IFN-β stimulus (N=7) during 8 and 24 hours. (A) sIFNAR2 expression in PBMC from each patient after 24 hours of culture in the presence or in the absence of IFN-β. (B) Boxplots comparing sIFNAR2 and MxA expression in unstimulated PBMC and IFN-β-stimulated PBMC during 8 and 24 hours of culture. Significant p values are marked with an asterisk. Logarithms are used to normalize and scale data for representation.

Journal: Frontiers in Immunology

Article Title: Soluble Receptor Isoform of IFN-Beta (sIFNAR2) in Multiple Sclerosis Patients and Their Association With the Clinical Response to IFN-Beta Treatment

doi: 10.3389/fimmu.2021.778204

Figure Lengend Snippet: In vitro study of IFNAR1, IFNAR2, sIFNAR2 and MxA gene expression. Representation of RNA relative expression of IFNAR1, IFNAR2, sIFNAR2 and MxA in PBMC from untreated MS patients cultured with or without IFN-β stimulus (N=7) during 8 and 24 hours. (A) sIFNAR2 expression in PBMC from each patient after 24 hours of culture in the presence or in the absence of IFN-β. (B) Boxplots comparing sIFNAR2 and MxA expression in unstimulated PBMC and IFN-β-stimulated PBMC during 8 and 24 hours of culture. Significant p values are marked with an asterisk. Logarithms are used to normalize and scale data for representation.

Article Snippet: Briefly, the plates were coated with 0.2-μg rabbit polyclonal anti-human IFNAR2 antibody (Abnova) overnight.

Techniques: In Vitro, Gene Expression, Expressing, Cell Culture