Journal: PLoS Pathogens

Article Title: Both α2,3- and α2,6-Linked Sialic Acids on O-Linked Glycoproteins Act as Functional Receptors for Porcine Sapovirus

doi: 10.1371/journal.ppat.1004172

Figure Lengend Snippet: LLC-PK cells were pretreated with 1 mM or 5 mM of NaIO 4 to remove carbohydrate moieties. (A) Alexa 594 or Alexa 594-labeled PSaV (MOI of 100 pfu/cell) were bound to pretreated LLC-PK cells, and were subsequently examined for bound virus by confocal microscopy. (B) 35 [S]Methionine/Cysteine-labeled PSaV, control FCV or CVB3 (50,000 c.p.m.) were bound to LLC-PK, CRFK or HeLa cells which were pretreated with or without NaIO 4 . Binding of radio-labeled PSaV, FCV or CVB3 was measured by liquid scintillation counting. (C) PSaV (MOI of 0.1 pfu/cell) was inoculated to NaIO 4 pretreated LLC-PK cells, and was subsequently analyzed by immunofluorescence assay to detect the viral antigen VPg, using a rabbit polyclonal antibody 72 h post infection. (D) PSaV, FCV or CVB3 positive cells (%) were quantified in three independent microscope fields. All experiments were performed three independent times, and figures A and C show one representative set of results. The scale bars correspond to 20 µm. Error bars indicate SD from triplicate samples. * p <0.05, ** p <0.005.

Article Snippet: The FCV F9 strain, human influenza A virus A/Puerto Rico/8/34 (H1N1) (PR8 virus) strain, human rotavirus DS-1 strain, and CVB3 Nancy strain were purchased from the ATCC.

Techniques: Labeling, Virus, Confocal Microscopy, Control, Binding Assay, Immunofluorescence, Infection, Microscopy

Journal: bioRxiv

Article Title: Effect of TSS on anti-CVB3 through TLR3 pathway

doi: 10.1101/2022.03.28.486160

Figure Lengend Snippet: The protective effect of drugs on the cardiac tissue of CVB3 virus-infected mice. a CVB3 mRNA levels in mouse hearts measured by qRT-PCR and normalized to GAPDH. TSS treatment can decrease the expression of CVB3 mRNA( ***P< 0.001 vs. model). b Cardiac index. ***P< 0.001 vs. model. c H&E staining of sectioned hearts (×400). Pathological changes were evaluated according to the structure of cardiac endothelial cells, myocardial fiber necrosis, calcium salt precipitation, and inflammatory cell infiltration. c1 normal group: the structure of epicardium and endocardial endothelial cells is complete and clear, the shape of myocardial fibers is relatively normal, no obvious degeneration and necrosis; c2 model group: local myocardial fibrosis and necrosis, epicardium and media layer myocardial fiber foci The necrosis and calcification were more obvious in the local area, and purple-black calcium salt deposition, a small amount of fibrous tissue and lymphocytes were seen in the necrotic area; c3 Ribavirin control group: myocarditis lesions were significantly alleviated, and only a small amount of myocardial fiber degeneration and necrosis; c4 TSS 66 mg·Kg -1 ·d -1 ; c5 TSS 132 mg·Kg -1 ·d -1 ; c6 TSS 264 mg·Kg -1 ·d -1 ; (TSS can significantly reduce the inflammatory lesions of myocardial tissue, and a small amount of myocardial fibrosis and necrosis)

Article Snippet: Coxsackievirus B3 strain (Nancy) was purchased from the China Center for Type Culture Collection.

Techniques: Virus, Infection, Quantitative RT-PCR, Expressing, Staining, Control

Journal: bioRxiv

Article Title: Effect of TSS on anti-CVB3 through TLR3 pathway

doi: 10.1101/2022.03.28.486160

Figure Lengend Snippet: TSS reduces the expression levels of TLR3, TRIF, TRAF6, IRF-3, NF-κB, MAPK, AP1 mRNA in the heart tissue of CVB3-infected Balb/c mice. TSS was added to 25 TCID50 CVB3 viral fluids to infect animals at a series of concentrations of 66, 132 and 264 mg/Kg. TLR3 and its downstream cytokine mRNA levels in each group were detected by qRT-PCR at d 6 after viral infection. *P< 0.05, **P< 0.01, ***P< 0.001 vs model; # P< 0.05, ## P< 0.01, ### P< 0.001 vs normal. In total, three independent experiments were performed.

Article Snippet: Coxsackievirus B3 strain (Nancy) was purchased from the China Center for Type Culture Collection.

Techniques: Expressing, Infection, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Effect of TSS on anti-CVB3 through TLR3 pathway

doi: 10.1101/2022.03.28.486160

Figure Lengend Snippet: Antiviral activity of TSS on CVB3 viruses in vitro. a-c Inhibition rates of TSS (from 2.4125 μM to 38.6451 μM) against CVB3. CVB3 with different infection protocols in HT-29 cells. Each concentration took up three wells for each assay, and three independent determinations were carried out, determined by CCK-8 assay. d CVB3 mRNA levels in HT-29 cells measured by qRT-PCR and normalized to GAPDH. TSS treatment can decrease the expression of CVB3 mRNA( ***P< 0.001 vs. model).

Article Snippet: Coxsackievirus B3 strain (Nancy) was purchased from the China Center for Type Culture Collection.

Techniques: Activity Assay, In Vitro, Inhibition, Infection, Concentration Assay, CCK-8 Assay, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Effect of TSS on anti-CVB3 through TLR3 pathway

doi: 10.1101/2022.03.28.486160

Figure Lengend Snippet: TSS reduces the expression levels of TLR3, TRIF, TRAF6, IRF-3, NF-κB, MAPK, AP1 in CVB3-infected HT-29 cells. TSS was added to 100 TCID50 CVB3 virus fluids to infect cells at a range of concentrations of 9.6604μM, 19.3226μM and 38.6451μM. The expression levels of TLR3 and its downstream cytokines in each group were detected by qRT-PCR and ELISA at 72h after virus infection. *P< 0.05, **P< 0.01, ***P< 0.001 vs model; # P< 0.05, ## P< 0.01, ### P< 0.001 vs normal. In total, three independent experiments were performed.

Article Snippet: Coxsackievirus B3 strain (Nancy) was purchased from the China Center for Type Culture Collection.

Techniques: Expressing, Infection, Virus, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay