Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: The cytotoxicity of Levistolide A on Vero and LLC-PK1 cells. Cells were treated with LA (20, 40, 60, 80, or 100 µM); DMSO served as the treatment control. The cell viability value was measured by CCK8 assay 48 h later. The value of each group was normalized by the average value (set at 100%) of the mock-treated group. ( A ) The cell viability of the Vero cells exposed to LA at designed concentrations. ( B ) The cell viability of LLC-PK1 cells exposed to LA at designed concentrations. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Control, CCK-8 Assay, Standard Deviation

Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: Levistolide A exhibits antiviral activity against PEDV strain DR13 and YN15 in Vero and LLC-PK1 cells. Vero cells were treated with LA at indicated concentrations and infected with PEDV at an MOI of 0.1 at the same time. DMSO served as the treatment control. Samples were collected at 36 hpi for IFA or TCID 50 assay analysis. ( A ) Immunofluorescence of PEDV-S protein (green) detected in PEDV strain DR13 infected Vero cells. ( B ) The viral titer determined in PEDV strain DR13 infected Vero cells. ( C ) Viral titer determined in PEDV strain YN15 infected Vero cells. ( D , E ) Vero cells were treated with LA (80 µM) or DMSO, and exposed to PEDV at 0.25 MOI or 0.5 MOI. Viral titers were assessed at 36 hpi. ( F , G ) Vero cells were treated with LA (80 µM) or DMSO, and exposed to PEDV (0.1 MOI). Viral titers were assessed at designed hpi. ( H ) Viral titer determined in the PEDV strain DR13 infected LLC-PK1 cells. ( I ) The viral titer determined in PEDV strain YN15 infected LLC-PK1 cells. Viral titer level was represented by the value of −lg TCID 50 /0.1 mL. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation. Asterisks indicate significant differences between mock-treated group and LA or DMSO treated group: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Activity Assay, Infection, Control, Immunofluorescence, Standard Deviation

Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: Levistolide A disturbed viral RNA and protein production in PEDV-infected cells. Vero cells were treated with LA at indicated concentrations and infected with PEDV at an MOI of 0.1 at the same time. DMSO served as the treatment control. The samples were collected at 36 hpi for TaqMan-based real-time RT-PCR or Western blot analysis. ( A ) Viral RNA copies determined in the PEDV strain DR13 infected cells; ( B ) viral RNA copies determined in PEDV strain CV777 infected cells; ( C ) viral RNA copies determined in PEDV strain YN15 infected cells. ( D ) The amount of PEDV N protein in samples was detected through Western blot; β-actin was set as the loading control. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation. Asterisks indicate significant differences between the mock-treated group and LA or DMSO treated group: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Infection, Control, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: Levistolide A exhibits an inhibitory effect on the PEDV-invading Vero cells. ( A ) Vero cells were exposed to PEDV (0.1 MOI) and treated with 80 µM LA or DMSO during 0–2 hpi, then the cultured supernatant was replaced by PEDV culture medium containing LA or DMSO at 2, 4, 6, 8 hpi. The cultured supernatant in all the groups was replaced by PEDV culture medium after 10 hpi and samples were collected at 36 hpi. ( B ) The viral titer determined in the PEDV strain DR13 infected Vero cells. ( C ) The viral titer determined in the PEDV strain YN15 infected Vero cells. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation. Asterisks indicate significant differences between virus control group and each LA treated group: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Cell Culture, Infection, Standard Deviation, Virus, Control

Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: Antiviral activity of Levistolide A against PEDV is associated with inducing ROS production. ( A ) The level of ROS in Vero cells or LLC-PK1 cells after incubation with LA (80 µM) for 24 h. The value of each group was normalized by the average value (set at 1) of the corresponding DMSO treated group. ( B ) Viral titer level in LA/NAC, LA, NAC, or DMSO treated group after exposing to PEDV for 36 h. Viral titer level was represented by value of − lg TCID50/0.1 mL. ( C ) Immunofluorescence of PEDV-S protein (green) detected in LA/NAC, LA, NAC, or DMSO treated group after exposing to PEDV for 36 h. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation. Asterisks indicate significant differences between virus control group and each LA treated group: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Activity Assay, Incubation, Immunofluorescence, Standard Deviation, Virus, Control

Journal: Viruses

Article Title: Levistolide A Inhibits PEDV Replication via Inducing ROS Generation

doi: 10.3390/v14020258

Figure Lengend Snippet: Levistolide A stimulated ER stress in Vero cells. ( A ) Tunicamycin (TM) potently inhibited PEDV replication. Vero cells exposed to PEDV strain YN15 (0.1 MOI) were incubated with 500 nM TM, samples were collected at 12, 24, and 36 hpi for TaqMan based real-time PCR; ( B ) mRNA level of ER stress-related genes increased in LA Vero cells. ( C ) The amount of GRP78 increased by treating with LA in either PEDV or mock infected Vero cells. PEDV strain YN15 (0.1 MOI) or mock infected Vero cells were incubated with LA or DMSO for 36 hpi. Western blot was performed to assess protein expression level and β-actin was set as loading control. Bands for GRP78 were marked by a black arrow. Data represent the mean ± standard deviation (SD) of three independent experiments, and error bars represent standard deviation. Asterisks indicate significant differences between virus control group and each LA treated group: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Levistolide A and Tunicamycin were purchased from MedChemExpress (Shanghai, China).

Techniques: Incubation, Real-time Polymerase Chain Reaction, Infection, Western Blot, Expressing, Control, Standard Deviation, Virus