Journal: Virology Journal

Article Title: Mechanism of HCV's resistance to IFN-α in cell culture involves expression of functional IFN-α receptor 1

doi: 10.1186/1743-422X-8-351

Figure Lengend Snippet: Western blot analysis of Jak-Stat signaling between S-5/15 and R-17/3 cell line . An equal number of S-5/15 and R-17/3 Huh-7 cells were treated with IFN-α 2 b (1000 IU/ml). Cells were harvested at different time intervals (in hrs) by trypsin-EDTA treatment, washed in PBS. Western blot analysis was performed using antibodies to IFNAR1, IFNAR2, pJak1, Jak1, pTyk2, Tyk2, pStat1, Stat1, pStat2, Stat2 and β-actin and then blots were developed by ECL kit. ( A) Expression level of Jak-Stat proteins between the two cell lines. Left panel shows the expression of Jak-Stat signaling proteins in the IFN-α sensitive Huh-7 cell line (S-5/15). Right panel shows the expression of Jak-Stat signaling proteins in the IFN-α resistant Huh-7 cell line (R-17/3). The level of β-actin protein in the blot indicated that equal amounts of protein lysate were loaded onto the gel in the Western analysis. (B) Show the expression of fully glycosylated mature form of IFNAR1 (~100 kD) and IFNAR2 (~90 kD) in all nine of different IFN-α resistant cell lines and one sensitive Huh-7 cell line (S-5/15) by Western blot analysis.

Article Snippet: The human IFNAR1 and three different IFNAR2 cDNA clones were purchased from OriGene Technologies (Rockville, MD, USA) and from our collaborator [ ].

Techniques: Western Blot, Expressing

Journal: Virology Journal

Article Title: Mechanism of HCV's resistance to IFN-α in cell culture involves expression of functional IFN-α receptor 1

doi: 10.1186/1743-422X-8-351

Figure Lengend Snippet: IFNAR1 protein expression complements the ISRE-firefly luciferase promoter activation in the resistant Huh-7 cells . The cured resistant Huh-7 cell line (R-17/3) was co-transfected with ISRE-firefly luciferase plasmid along with each expression plasmid by FuGene6 transfection reagent. The pRL-TK plasmid was used as a transfection control. After this step, cells were treated with IFN-α (1000 IU/ml). After 24 hours, cells were lyzed and luciferase activity was measured. The graphs represent a relative to control luciferase activity. ( A) The activation of ISRE promoter by IFN-α in R-17/3 cells expressing different proteins of Jak-Stat signaling expressed as relative luciferase units (RLU) between IFN-α treated and untreated. Expression of wild type full-length IFNAR1 protein is able to induce the ISRE-Luciferase activity. ( B) The induction of ISRE-luciferase activity in the R-17/3 cells by IFN-α in the presence of three different variants of IFNAR2 and wild type IFNAR1 was depicted as fold increase as compared to untreated cells. ( C) IFN-α induces the ISRE-Firefly luciferase activity (fold increase) in all nine IFN-α resistant Huh-7 cell lines when they were co-transfected with the wild type IFNAR1 plasmid.

Article Snippet: The human IFNAR1 and three different IFNAR2 cDNA clones were purchased from OriGene Technologies (Rockville, MD, USA) and from our collaborator [ ].

Techniques: Expressing, Luciferase, Activation Assay, Transfection, Plasmid Preparation, Control, Activity Assay

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: Summary of genome-wide significant pQTLs.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques: Genome Wide

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: (A-C) Data on IFNAR2 cell surface levels. (A) Regional association plot for IFNAR2 surface levels in CD56 dim NK cell and (B) boxplots stratified by rs28488669 and rs2186355. (C) Heat-map of the relation between p-values for the two cis -IFNAR2 pQTLs in different cell-types as specified. (D-E) Data on IFN-α induced phosphorylation of STAT4 (pSTAT4) and STAT1 (pSTAT1). (D) Regional association plot of the IFN-α induced pSTAT4 in CD56 dim NK cells and (E) boxplots stratified by rs2298260. (F) Heat-map of the relation between pQTL p-values for IFN-α induced pSTAT1 and pSTAT4 in indicated cell-types. (A and D) The genome-wide significance level (p < 5.0E-8) is denoted by a red line. (A-F) p-values from the full model using single (A, D-F), or two additive SNPs (B-C), ( n = 303). Boxplots show median, IQR and range. gMFI = geometric mean fluorescence intensity.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques: Phospho-proteomics, Genome Wide, Fluorescence

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: (A) Regional association plots of the trans -IFNAR2 pQTL in B cells before (top) and after conditioning for rs2735099 (middle) and rs17199328 (bottom). (B-D) Boxplots of IFNAR2 levels in B cells (B), CD8 + T cells (C) and CD4 + T (D) cells stratified by rs2735099 and rs17199328 ( n = 301). (E-F) IFNAR1 (E) and IFNGR2 (F) surface levels in B cells ( n = 95). p-values from the full model using single (A, E and F), or two additive SNPs (B-D). Boxplots show median, IQR and range. gMFI = geometric mean fluorescence intensity.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques: Fluorescence

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: (A) Correlation between rs2735099 and rs17199328 genotypes and imputed HLA-A ( n = 301) and HLA-B alleles ( n = 303). (B) The proportion of B cells subsets in bulk B cells ( n = 95). (C) IFNAR2 surface levels in subsets of B cells from healthy donors ( n = 95). (D) IFNAR2 surface levels in subsets of B cells and T cells from healthy donors ( n = 22) and PwMS ( n = 26). (B-D) Data are stratified by the sum of the MS-protective class I alleles HLA-A*02, HLA-A*68 and HLA-B*44, as indicated. p-values from the full model (B-C) or a simple linear regression (D), using the sum of MS-protective alleles (0–3) as a continuous variable. Boxplots show median, IQR and range. gMFI = geometric mean fluorescence intensity.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques: Fluorescence

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: (A) A schematic picture of the IFNAR2 isoforms. (B) IFNAR2bc , IFNAR2b and IFNAR2c mRNA levels in isolated B cells determined using qRT-PCR. mRNA levels are normalized to the expression of the reference gene RPL13A and expressed as arbitrary units (AU) ( n = 47, simple linear regression using the sum of MS-protective class I alleles (HLAsum) as independent variable). (C) Correlation between IFNAR2bc mRNA levels and IFNAR2 surface receptor levels determined using flow cytometry ( n = 47). Data are binned into 0, 1 and ≥2 HLAsum. p-values and Pearson’s correlation coefficient (r) from linear regressions of each group are shown in coloured text. p-values from multiple linear regressions of the combined data using HLAsum (continuous variable) and mRNA values as independent variables in the full model are denoted in black. (D) Heat-maps comparing the correlation between IFNAR2 levels and IFN-α induced pSTAT1 (top) or pSTAT4 (bottom) in indicated cell-types with and without including HLAsum as a covariate in the full model ( n = 303). (E-G) Correlation between IFNAR2 protein levels and IFN-α (2000 IU/ml) induced pSTAT1 (E), pSTAT4 (F) and CXCL10 (G) in B cells. p-values as described in (C), ( n = 303). Boxplots show median, IQR and range. gMFI = geometric mean fluorescence intensity.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques: Isolation, Quantitative RT-PCR, Expressing, Flow Cytometry, Fluorescence

Journal: PLoS Genetics

Article Title: Function of multiple sclerosis-protective HLA class I alleles revealed by genome-wide protein-quantitative trait loci mapping of interferon signalling

doi: 10.1371/journal.pgen.1009199

Figure Lengend Snippet: (A) Cis -eQTL analysis within ± 1 Mb from the HLA-A*02/A*68 and HLA-B*44 tag SNPs in LCLs ( n = 373). The p-value threshold of p < 3.2E-4, corresponding to a Bonferroni adjusted p-value of < 0.05 (155 analysed genes) is denoted with dotted lines. Data for 28 of the 155 genes are shown. The inset shows the transcript specific cis -eQTL p-values of HLA-J . (B) Correlation between p-values from the IFNAR2 pQTL in B cells and the cis -eQTL in LCLs for the transcript HLA-J-001 (left) and HCG4P5-001 (right). The HLA-A*02/A*68 and the B*44 tagging SNPs are marked in red and blue respectively. (C) Cis -meQTL analysis of the HLA-A*02/A*68 tagging SNP rs2735099 in primary B cells ( n = 43). The p-value threshold of p < 2.1E-6, corresponding to a Bonferroni adjusted p-value of < 0.05 (23, 722 analysed CpG-sites) is denoted with dotted red lines.

Article Snippet: The IFNAR2 mAb REA124 was validated by transfection of HEK293T cells with siRNA targeting IFNAR2 ((dTdT-)GCACCATAGTGACACTGAA-dTdT), and plasmids encoding IFNAR2b or IFNAR2c (#RC201212 and #RC238664, Origene) ( ).

Techniques:

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

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

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The primers involved in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: The antibodies used in this study.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques:

Journal: Poultry Science

Article Title: New insights into the spleen injury by mitochondrial dysfunction of chicken under polystyrene microplastics stress

doi: 10.1016/j.psj.2024.103674

Figure Lengend Snippet: MPs induced ferroptosis in spleen. (A) Western Blot results in ferroptosis-related proteins. (B) Quantitative analysis of GPX4, FTH1, and SLC7A11 protein expression. (C) Ferroptosis-related factors of mRNA expression changes. Data were expressed as mean ± SD, n = 3. *, **, ***, denotes: p < 0.05, 0.01, and 0.001, respectively.

Article Snippet: GPX4 , 1:500 , Boster Biotechnology, China , A02059-1.

Techniques: Western Blot, Expressing