Journal: Frontiers in Immunology

Article Title: HSP90AA1 interacts with CSFV NS5A protein and regulates CSFV replication via the JAK/STAT and NF-κB signaling pathway

doi: 10.3389/fimmu.2022.1031868

Figure Lengend Snippet: HSP90AA1 over-expression activates STAT1 and P65 in PK-15 and 3D4/2 cells. (A) HSP90AA1 over-expression affected the phosphorylation of STAT1 and nuclear translocation of p-STAT1 in PK-15 and 3D4/2 cells were observed by laser confocal microscopy. (B) HSP90AA1 over-expression affected the nuclear translocation of p-P65 in PK-15 and 3D4/2 cells were observed by laser confocal microscopy.

Article Snippet: Primary antibodies include rabbit anti-p-STAT1 pAb (Bioss, bs-3427R), rabbit-anti-p-P65 pAb (Affinity Biosciences, AF2006) and mouse anti-Flag mAb.

Techniques: Over Expression, Translocation Assay, Confocal Microscopy

Journal: Frontiers in Immunology

Article Title: HSP90AA1 interacts with CSFV NS5A protein and regulates CSFV replication via the JAK/STAT and NF-κB signaling pathway

doi: 10.3389/fimmu.2022.1031868

Figure Lengend Snippet: CSFV infection antagonizes the activation of HSP90AA1 on JAK/STAT pathway. (A, B) Western blot for JAK1, STAT1, p-JAK1, p-STAT1, OAS2, ISG-15 and HSP90AA1 expression in HSP90AA1-overexpression or knock-downed PK-15 and 3D4/2 cells. Cells were infected with CSFV (MOI=1) after transfection p3×Flag-HSP90AA1 or siHSP90AA1. The cells were not infected with CSFV as a control. Cells were harvested at 24hpi and 48hpi served to Western blot. The relative levels of proteins were estimated by histograms representing density reading of the gel bands with Image J, and the ratios were calculated relative to tubulin control. (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 calculated using two-way ANOVA, ns, not significant).

Article Snippet: Primary antibodies include rabbit anti-p-STAT1 pAb (Bioss, bs-3427R), rabbit-anti-p-P65 pAb (Affinity Biosciences, AF2006) and mouse anti-Flag mAb.

Techniques: Infection, Activation Assay, Western Blot, Expressing, Over Expression, Transfection

Journal: Journal of Veterinary Science

Article Title: Role of IFNLR1 gene in PRRSV infection of PAM cells

doi: 10.4142/jvs.2021.22.e39

Figure Lengend Snippet: PAMs were transfected with IFNLR1 RNAi and the overexpression plasmid. Total cellular proteins were extracted, and the protein expressions of p-STAT1 and STAT1 were detected by performing western blot analysis.

Article Snippet: Antibody against STAT1 (bs-9584R rabbit anti-STAT1 antibody) was purchased from Bioss Biotech (Beijing, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Western Blot

Journal: Journal of Veterinary Science

Article Title: Role of IFNLR1 gene in PRRSV infection of PAM cells

doi: 10.4142/jvs.2021.22.e39

Figure Lengend Snippet: PAMs signal via the JAK/STAT pathway. (A) JAKI was added to PAMs transfected with the pcDNA3.1(+) plasmid or pcDNA3.1(+)-IFNLR1 plasmid for 1 h and then infected with PRRSV for 48 h. The phosphorylation level of STAT1 was detected by performing western blot analysis. (B) The CCK8 assay was used to detect changes in PAM proliferation ability after adding JAKI. Data represent mean ± SD values of at least three independent experiments. Significant differences are indicated with different letters, etc., a, b, c as determined by the Duncan's multiple range test ( p < 0.05).

Article Snippet: Antibody against STAT1 (bs-9584R rabbit anti-STAT1 antibody) was purchased from Bioss Biotech (Beijing, China).

Techniques: Transfection, Plasmid Preparation, Infection, Western Blot, CCK-8 Assay