ev d68  (ATCC)

Journal: Genes & Diseases

Article Title: Salt-inducible kinase 1 is a key gene in suppressing EVD68-induced asthma by modulating antiviral immunity

doi: 10.1016/j.gendis.2025.101845

Figure Lengend Snippet: The common differentially expressed genes between EV-D68 infection and asthma. (A, B) Volcano plot of differentially expressed genes (DEGs) in the EV-D68-related GSE184488 dataset and asthma-related GSE143303 dataset. (C) Common DEGs in the GSE184488 dataset and the GSE143303 dataset were represented by Venn diagrams. (D) KEGG enrichment analysis of the 74 common DEGs identified in EVD68 infection and asthma. (E) GO enrichment analysis of the 74 common DEGs identified in EVD68 infection and asthma. BP, biological process; CC, cellular component; MF, molecular function.

Article Snippet: The EV-D68 (ATCC VR-1826), EV-A71, HSV-1, and VSV-GFP were kept in our laboratory.

Techniques: Infection

Journal: Genes & Diseases

Article Title: Salt-inducible kinase 1 is a key gene in suppressing EVD68-induced asthma by modulating antiviral immunity

doi: 10.1016/j.gendis.2025.101845

Figure Lengend Snippet: SIK1 expression is induced by EV-D68 infection in vivo . Eight-to-ten-week-old type I interferon receptor-deficient mice ( Ifna −/− ) were treated with EV-D68 (50 μL of 1 × 10 7 PFU/mL viral stock for each mouse) or an equal volume of phosphate-buffered saline through the intranasal route after anesthesia. Lungs were harvested 48 h after infection and then analyzed by hematoxylin-eosin staining, quantitative PCR, and western blotting. (A) Images showing lung inflammation 48 h after treatment with EV-D68 or mock phosphate-buffered saline, as visualized by hematoxylin-eosin staining (scale bar: 50 mm). (B–I) The indicated genes were detected by quantitative PCR and normalized to GAPDH expression. Values are from three independent experiments and expressed as mean ± standard deviation. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (J) The protein expression level of SIK1 in the lungs of mice from the mock and EV-D68 groups was detected by western blotting.

Article Snippet: The EV-D68 (ATCC VR-1826), EV-A71, HSV-1, and VSV-GFP were kept in our laboratory.

Techniques: Expressing, Infection, In Vivo, Saline, Staining, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation

Journal: Genes & Diseases

Article Title: Salt-inducible kinase 1 is a key gene in suppressing EVD68-induced asthma by modulating antiviral immunity

doi: 10.1016/j.gendis.2025.101845

Figure Lengend Snippet: SIK1 shows antiviral effects in various viral infections. (A, B) A549 cells were infected with EV-D68 (MOI = 0.1 or 1) for 24 h. (A) Quantitative reverse transcription PCR (RT-qPCR) analysis of relative SIK1 mRNA expression. The results were normalized to GAPDH expression. (B) Western blotting analysis of SIK1 and EV-D68 VP1 protein expression. β-actin was used as the loading control. Values were from three independent experiments and expressed as mean ± standard deviation. (C, D) RD cells were infected with EV-A71 (MOI = 0.1 or 0.5) for 24 h. (C) RT-qPCR analysis of relative SIK1 mRNA expression. The results were normalized to GAPDH expression. (D) Western blotting analysis of SIK1 and EV-A71 VP1 protein expression. β-actin was used as the loading control. Values were from three independent experiments and expressed as mean ± standard deviation. (E, F) A549 cells were transfected with si-SIK1 or si-NC and then infected with EV-D68 (MOI = 0.5) for 24 h. (E) The relative viral RNA copy numbers were determined by RT-qPCR and normalized to GAPDH. (F) The protein expression levels of EV-D68 VP1 and SIK1 were detected by western blotting. β-actin was used as the loading control. Values were from three independent experiments and expressed as mean ± standard deviation. (G, H) A549 cells were transfected with plasmid pCDH-SIK1 or empty pCDH vector and then infected with EV-D68 (MOI = 0.5) for 24 h. The viral replication and protein expression level of EV-D68 VP1 and SIK1 were detected as described above. Values were from three independent experiments and expressed as mean ± standard deviation. (I, J) A549 cells were infected with VSV-GFP for 6 h (MOI = 0.1, 0.5) or HSV-1 for 24 h (MOI = 0.1, 0.2), and then the relative mRNA expression of SIK1 was analyzed by RT-qPCR. Values were from three independent experiments and expressed as mean ± standard deviation. (K, L) Box plots represent the normalized expression levels of SIK1 using Z-score normalization in GSE157103 (for CV-A6) and GSE243200 (for SARS-COV-2) datasets. SIK1 expression correlation was analyzed using Spearman's method. (M, N) A549 cells were transfected with si-SIK1 or si-NC and then infected with VSV-GFP (MOI = 0.5) for 12 h. The replication of VSV-GFP was visualized by immunofluorescence microscopy (scale bar: 50 μm), and VSV-GFP RNA synthesis was determined by RT-qPCR analysis. Values were from three independent experiments and expressed as mean ± standard deviation. (O, P) A549 cells were transfected with plasmid pCDH-SIK1 or empty pCDH vector and then infected with VSV-GFP (MOI = 0.5) for 12 h. To detect viral replication by quantitative PCR, viral titers were determined by plaque assay as described in the methods section, and viral RNA synthesis was determined by RT-qPCR analysis. Values were from three independent experiments and expressed as mean ± standard deviation. (Q, R) SIK1 was interfered with by si-SIK1 or overexpressed via transfection of plasmid pCDH-SIK1 in A549 cells, and then the cells were infected with HSV-1. Viral replication was determined by RT-qPCR. Values were from three independent experiments and expressed as mean ± standard deviation. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, non-significant.

Article Snippet: The EV-D68 (ATCC VR-1826), EV-A71, HSV-1, and VSV-GFP were kept in our laboratory.

Techniques: Infection, Reverse Transcription, Quantitative RT-PCR, Expressing, Western Blot, Control, Standard Deviation, Transfection, Plasmid Preparation, Immunofluorescence, Microscopy, Real-time Polymerase Chain Reaction, Plaque Assay

Journal: Genes & Diseases

Article Title: Salt-inducible kinase 1 is a key gene in suppressing EVD68-induced asthma by modulating antiviral immunity

doi: 10.1016/j.gendis.2025.101845

Figure Lengend Snippet: Metformin-mediated activation of SIK1 protects against EV-D68-driven asthma exacerbation in house dust mite (HDM)-sensitized mice. (A) C57BL/6 mice (6–8 weeks) were administered metformin at doses of 100 mg/kg or 250 mg/kg once daily via intraperitoneal injection on day 1 and day 2. On day 3, lung tissues were collected, and the protein level of SIK1 was determined by western blotting analysis. (B) Experimental timeline. C57BL/6 mice (6–8 weeks) were intranasally sensitized with 250 μg kg −1 HDM extract on day 0 and challenged daily with the same dose on days 7–11. On days 12–13, animals received EV-D68 (1 × 10 6 PFU/kg) or DMEM (vehicle) intranasally. Metformin (100 mg/kg, intraperitoneal) was administered once daily on days 12–14. Airway hyper-responsiveness measurements and broncho-alveolar lavage fluid (BALF) collection were performed on day 15; lung tissue was used for quantitative PCR analyses. (C) Airway responsiveness to increasing doses of methacholine. (D) Differential cell counts of BALF by Wright-Giemsa staining. (E – H) The indicated genes were detected by quantitative PCR and normalized to GAPDH expression. Values were from three independent experiments and expressed as mean ± standard deviation. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: The EV-D68 (ATCC VR-1826), EV-A71, HSV-1, and VSV-GFP were kept in our laboratory.

Techniques: Activation Assay, Injection, Western Blot, Real-time Polymerase Chain Reaction, Staining, Expressing, Standard Deviation