non-structural protein 1 (ns1, gtx638102) antibody  (GeneTex)

Journal: Heliyon

Article Title: Generation of rescued Japanese encephalitis virus genotype 1 from infectious full-size clone using reverse genetics

doi: 10.1016/j.heliyon.2024.e33142

Figure Lengend Snippet: Expression of JEV E and NS1 proteins. Vero cells were infected with rJEV-GI or-K05GS at an MOI of 0.1, and the cell culture supernatant and lysates were harvested at 5 dpi. (A) SDS-PAGE followed by Coomassie Brilliant Blue G-250 staining (B) Western blot analysis with specific antibodies against JEV E and NS1 protein. M, protein size marker; 1: mock-infected cell lysate; 2: rJEV-GI-infected cell lysate; 3: K05GS-infected cell lysate; 4: mock-infected cell supernatant; 5: rJEV-GI-infected cell supernatant; 6: K05GS-infected cell supernatant. Asterisks indicate a possible cleavage fragment of the JEV NS1 protein (50 kDa) following the instructions from Abcam (ab41651). This experiment was repeated twice, and no significant differences were found. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Anti -JEV envelope (E) antibody (GeneTex; 1:1000 dilution) and anti-JEV non-structural protein 1 (NS1) antibody (Abcam; 1:1000 dilution) were reacted with the membrane.

Techniques: Expressing, Infection, Cell Culture, SDS Page, Staining, Western Blot, Marker

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Increase in viral RNA and protein levels by LPCs. ( A ) Relative expression levels of IAV genes in THP-1 cells treated with different dosages of LPCs. Color boxes indicated control (□), IAV (■) and IAV with LPC (▒). ( B ) The protein level of IAV proteins in THP-1 cells treated with LPCs. RNA levels were determined using RT-qPCR, and protein expression was detected using Western blotting. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001, ** p < 0.005, * p < 0.05 compared with the values obtained for cells infected with IAV. M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Infection

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Differential expression of viral RNAs according to the type of LPCs. The relative expression levels of IAV genes were determined using RT-qPCR. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001, ** p < 0.005, * p < 0.05 compared with the values obtained for cells infected with IAV. Color boxes indicated control (□), IAV (■) and IAV with LPCs (▒). M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Quantitative Proteomics, Expressing, Quantitative RT-PCR, Infection, Control

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Attenuation of viral gene expression by MAP kinase inhibitor (SB203580), JNK inhibitor, and PI3K inhibitor (LY294002) in LPC-treated THP-1 cells. Relative expression levels of IAV genes were determined using RT-qPCR. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001 compared with values obtained for cells infected with IAV, # p < 0.05 compared with values obtained for cells infected with IAV and treated with LPC. Color boxes indicated control (□), IAV (■) and IAV with LPC and inhibitors (▒). M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Infection, Control

Journal: Viruses

Article Title: The Interferon-Induced Protein with Tetratricopeptide Repeats Repress Influenza Virus Infection by Inhibiting Viral RNA Synthesis.

doi: 10.3390/v15071412

Figure Lengend Snippet: Figure 2. Effects of IFIT1 or IFIT2 knockdown on influenza virus infection in lung epithelial A549 cells. A549 cells were transduced with lentiviral shRNA control (shCon), shIFIT1 or shIFIT2 at an MOI of 100 for 48 h and then infected with PR/8 at an MOI of 0.01 for 48 h. (A) The representative western blots. (B–D) Quantitation of IFIT1, IFIT2, viral NP and NS1. (C) Virus titers from culture media. Data shown are means ± SE. ** p < 0.01, *** p < 0.001, and **** p < 0.0001. n = 3. One-way (C–E) and two-way ANOVA (B), followed by the Tukey test.

Article Snippet: Western blot analysis was performed using the following primary antibodies and dilutions: mouse anti-NP (HB-65, ATCC, 1:50), mouse anti-non-structural protein 1 (NS1) (#sc-130568, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), mouse anti-βactin (Thermo Fisher Scientific, 1:3000), goat anti-IFIT1 (#PA5-848, Invitrogen, 1:1000), and mouse anti-IFIT2 (#sc-390724, Santa Cruz, 1:1000).

Techniques: Knockdown, Virus, Infection, Transduction, shRNA, Control, Western Blot, Quantitation Assay

Journal: Viruses

Article Title: The Interferon-Induced Protein with Tetratricopeptide Repeats Repress Influenza Virus Infection by Inhibiting Viral RNA Synthesis.

doi: 10.3390/v15071412

Figure Lengend Snippet: Figure 3. Effects of IFIT1 or IFIT2 overexpression on influenza infection in HEK293 cells: (A–D) HEK293 cells were transfected with a vector control, Flag-tagged IFIT1 or IFIT2 expres- sion plasmid (IFIT1-OE and IFIT2 OE) for 48 h and then infected with PR/8 at an MOI of 0.01 for 48 h (A–D) or at an MOI of 5 for 16 h (E,F). (A) IFIT mRNA levels; (B) representative Western blots; (C) quantitation of viral NP and NS1 protein levels; (D) viral NP and NS1 RNA levels; (E) immunoflu- orescence staining of HEK293 cells with ant-NP antibodies; (F) Percentage of NP-positive cells. Data shown are means ± SE. * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3. Two-way ANOVA (A,C,D) and one-way ANOVA (F), followed by the Tukey test.

Article Snippet: Western blot analysis was performed using the following primary antibodies and dilutions: mouse anti-NP (HB-65, ATCC, 1:50), mouse anti-non-structural protein 1 (NS1) (#sc-130568, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), mouse anti-βactin (Thermo Fisher Scientific, 1:3000), goat anti-IFIT1 (#PA5-848, Invitrogen, 1:1000), and mouse anti-IFIT2 (#sc-390724, Santa Cruz, 1:1000).

Techniques: Over Expression, Infection, Transfection, Plasmid Preparation, Control, Western Blot, Quantitation Assay, Staining

Journal: Viruses

Article Title: The Interferon-Induced Protein with Tetratricopeptide Repeats Repress Influenza Virus Infection by Inhibiting Viral RNA Synthesis.

doi: 10.3390/v15071412

Figure Lengend Snippet: Figure 5. Effects of IFIT1 or IFIT2 knockdown or overexpression on viral RNA synthesis and polymerase activity: (A,B) Viral RNA synthesis. A549 cells were transduced with lentiviral shRNA control (shCon), shIFIT1 or shIFIT2 at an MOI of 100 for 48 h and then infected with PR/8 at an MOI of 5 for 5 h. NP and NS1 viral RNA (vRNA, cRNA and mRNA) levels were determined. (C,D) Polymerase activity. HEK293 cells were transfected with shRNA control (shCon), shIFIT1, or shIFIT2 (C) or vector control, Flag-tagged IFIT1 or IFIT2 expression plasmid (IFIT1-OE and IFIT2 OE) (D) for 24 h, and then co-transfected with minigenome vectors, an IAV luciferase reporter vector and pRL-TK normalization vector for another 24 h. Dual-luciferase activities were determined. Polymerase activity was expressed by the ratio of Firefly to Renilla activities and then normalized to shCon (C) or vector control (D). Data shown are means ± SE. * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3. Two-way ANOVA (A,D) and one-way ANOVA (C,D), followed by the Tukey test.

Article Snippet: Western blot analysis was performed using the following primary antibodies and dilutions: mouse anti-NP (HB-65, ATCC, 1:50), mouse anti-non-structural protein 1 (NS1) (#sc-130568, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), mouse anti-βactin (Thermo Fisher Scientific, 1:3000), goat anti-IFIT1 (#PA5-848, Invitrogen, 1:1000), and mouse anti-IFIT2 (#sc-390724, Santa Cruz, 1:1000).

Techniques: Knockdown, Over Expression, Activity Assay, Transduction, shRNA, Control, Infection, Transfection, Plasmid Preparation, Expressing, Luciferase

Journal: One Health

Article Title: Assessing West Nile virus (WNV) and Usutu virus (USUV) exposure in bird ringers in the Netherlands: a high-risk group for WNV and USUV infection?

doi: 10.1016/j.onehlt.2023.100533

Figure Lengend Snippet: .IgG protein microarray signals of bird ringers and control groups. WNV and USUV NS1 protein microarray IgG titers (log2 scale) for A) Bird ringers (N = 157, 2021), B) Health care workers (negative panel, N = 58, 2009–2010), C) Dutch general population survey (PIENTER, N = 94, 2016–2017), * and ° symbols indicate the same participants, and D) Dutch blood donors (Sanquin, N = 96, 2021).

Article Snippet: Slides were printed with WNV (Sino Biological) and USUV non-structural proteins 1 (NS1) (The Native Antigen company) proteins as well as JEV and TBEV NS1 (Immune Technology) to assess flavivirus antibody cross-reactivity.

Techniques: Microarray