mouse anti non structural protein 1 ns1  (Santa Cruz Biotechnology)

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: Scientific Reports

Article Title: Caveolae-mediated albumin transcytosis is enhanced in dengue-infected human endothelial cells: A model of vascular leakage in dengue hemorrhagic fever

doi: 10.1038/srep31855

Figure Lengend Snippet: (A) Intracellular BSA in DENV2-infected EA.hy926 cells at various post-infection time-points was determined by Western blot analysis. GAPDH served as the loading control while non-structural 1 (NS1) was a marker for DENV infection. (B) Quantitative analysis of BSA band intensity (C) Detection of BSA conjugated with Alexa Fluor 488 (Alexa488-BSA) (shown in green) by a laser scanning confocal microscope (LSM 510 Meta, Carl Zeiss). Hoechst staining was performed to demonstrate nuclei. DENV stained in red served as the marker for successful DENV2 infection. Original magnification was 630X for all panels. (D) Fluorescence intensity of the internalized Alexa488-BSA was quantitated using ImageJ software. (n = 3 independent experiments for both Western blot analysis and fluorescence study; * p < 0.05 vs. mock control; # p < 0.01 vs. mock control).

Article Snippet: A mouse monoclonal antibody against DENV non-structural protein 1 (NS1) (Abcam; Cambridge, UK) was also used to detect NS1 as a marker for DENV infection.

Techniques: Infection, Western Blot, Control, Marker, Microscopy, Staining, Fluorescence, Software