Journal: Psychiatry and Clinical Psychopharmacology

Article Title: Antinuclear Antibodies and Thyroid Autoantibodies in the Serum of Chinese Patients with Acute Psychiatric Disorders: A Retrospective Study

doi: 10.5152/pcp.2022.22005

Figure Lengend Snippet: Association of the Thyroid Autoantibody Status with ANA

Article Snippet: Total (n = 834) , 73+/115− , 167+/479− , Pearson’s Chi square test , .001.

Techniques: Control

Journal: Journal of Virology

Article Title: The Andes Orthohantavirus NSs Protein Antagonizes the Type I Interferon Response by Inhibiting MAVS Signaling

doi: 10.1128/JVI.00454-20

Figure Lengend Snippet: The ANDV-NSs protein suppresses the IFN induction pathway downstream of MDA5 and RIG-I and upstream of TBK1. (A) Schematic representation of IFN-β–Fluc, SV40-Rluc, pHA-EV, pCAGGs-FLAG-EV, or pCAGGs-FLAG encoding the FLAG versions of MDA5, RIG-I-2CARD, MAVS, TBK1, or IRF3-5D plasmids used in the assays. (B) HEK-293T cells were transfected with IFN-β–Fluc, SV40-Rluc, pHA-EV, pCAGGs-FLAG-EV, or pCAGGs-FLAG encoding the FLAG versions of MDA5, RIG-I-2CARD, MAVS, TBK1, or IRF3-5D, all inducers of the IFN induction pathway by polyethyleneimine (PEI). At 18 h p.t., the cells were lysed, and FLuc and RLuc luciferase activities were determined using a commercial dual-luciferase assay (Promega). Values represent the relative luciferase activity (FLuc/RLuc) obtained for the IFN-β–FLuc reporter plasmids when transfected with the inducer plasmids over the FLuc/RLuc activity obtained when transfecting the FLAG-EV plasmid, which was arbitrary set as 1.Values shown are the mean (±SEM) of at least three independent experiments, each conducted in triplicate. Statistical analysis was performed with a Student’s t test (*, P < 0.05). HEK 293T cells were transfected with plasmids expressing the HA-tagged ANDV-NSs, IAV-NS1, and SNV-N proteins or with HA-EV along with plasmids expressing the FLAG-tagged version of MDA5 (C) RIG-I (D), TBK1 (E), or IRF-5D (F) and the plasmids IFN-β–FLuc and SV40-RLuc. At 18 h p.t., the cells were lysed with passive lysis buffer (Promega), and FLuc and RLuc activities were determined using a dual-luciferase assay (Promega). The relative luciferase activity (FLuc/RLuc) obtained for IFN-β–FLuc when transfected with the plasmid pHA-EV together with the inducer of the IFN production pathway was set to 100%. The values shown are the mean (±SEM) of at least three independent experiments, each conducted in triplicate. Statistical analysis was performed by ANOVA followed by Dunnett’s multiple-comparison test (*, P < 0.05). A representative Western blot for each set of experiments is shown below its corresponding bar chart.

Article Snippet: Total protein (1 mg) was used for immunoprecipitation assays together with protein A/G Plus agarose (sc-2003; Santa Cruz Biotechnology) loaded with anti-HA mouse monoclonal antibody (H9658; Sigma-Aldrich), anti-MAVS mouse monoclonal antibody (sc-166583; Santa Cruz Biotechnology), or anti-TBK1 mouse monoclonal antibody (sc-73115; Santa Cruz Biotechnology).

Techniques: Transfection, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Lysis, Western Blot

Journal: Journal of Virology

Article Title: The Andes Orthohantavirus NSs Protein Antagonizes the Type I Interferon Response by Inhibiting MAVS Signaling

doi: 10.1128/JVI.00454-20

Figure Lengend Snippet: The HA-NSs protein and Flag-MAVS bind in HEK 293T cells. HEK293T cells were cotransfected with Flag-MAVS, Flag-TBK1, and HA-EV or HA-NSs plasmids. At 24 h p.t., cells were lysed, and immunoprecipitation (IP) assays were performed using protein A/G agarose coated with the corresponding antibody. The beads were washed extensively 4 times and incubated with loading buffer at 95°C to rescue the supernatant, which was used for Western blotting. (A) Whole-cell lysate (WCL) of each sample and (B) IP fractions with the corresponding primary antibody highlighted on the left side of the panels. Western blotting was performed using mouse anti-TBK1, mouse anti-MAVS, mouse anti-HA, and mouse anti-GAPDH. horseradish peroxidase (HRP)-conjugated protein A/G was used to detect the primary antibodies.

Article Snippet: Total protein (1 mg) was used for immunoprecipitation assays together with protein A/G Plus agarose (sc-2003; Santa Cruz Biotechnology) loaded with anti-HA mouse monoclonal antibody (H9658; Sigma-Aldrich), anti-MAVS mouse monoclonal antibody (sc-166583; Santa Cruz Biotechnology), or anti-TBK1 mouse monoclonal antibody (sc-73115; Santa Cruz Biotechnology).

Techniques: Immunoprecipitation, Incubation, Western Blot