DOI: 10.57760/sciencedb.28396
title: A dataset of Verapamil HCl suppresses PRRSV infection
title: A dataset of Verapamil HCl suppresses PRRSV infection

description: Raw data for Figure1B derived from Cytotoxicity of Verapamil HCl against MARC-145 cells, PK-15CD163 cells, and PAMs, these cells were treated with different doses of Verapamil HCl (10, 20, 40, 60, 80, and 100 µM) for 24 h, cell viability was detected using the CCK-8 kit. The viable cell count was assessed through absorbance measurements at 450 nm using a microplate reader. In relative cell viability calculations, the untreated cells served as blank controls.Raw data for Figure1D, F, H derived from Verapamil HCl suppresses PRRSV SD16 infection in a dose-dependent manner. MARC-145 cells, PK-15CD163 cells, and PAMs were infected with PRRSV SD16 (MOI = 0.1) pre-incubated with Verapamil HCl (5, 10, 20, 40, and 80 µM). progeny virus copies was calculated using standard curve. The data shown are representative of three independent experiments and were analyzed using Student’s t-tests. ** p &lt; 0.01 and *** p &lt; 0.001: compared with 0 µM Verapamil HCl-treated cells. Raw data for Figure2 derived from anti-PRRSV effects of Verapamil HCl. MARC-145 cells and PAMs were incubated with 0µM or 80 µM Verapamil HCl for 2 h. The cells were infected with various PRRSV-1 and PRRSV-2 strains at 0.1 MOI for 1 h at 37°C. At 24 hpi, the cells were harvested to measure PRRSV N protein expression at the transcription and protein levels using qPCR, GAPDH served as the reference gene, the relative quantification of ORF-7 expression was performed using the 2−∆∆Ct method. Raw data for Figure3 derived from the stage of Verapamil HCl action during the PRRSV life cycle. A, PRRSV SD16 was pre-incubated with Verapamil HCl (80 µM) or DMSO for 2 h at 37°C. Then, this mixture was inoculated into MARC-145 cells. progeny virus copies was calculated using standard curve to evaluate the virucidal activity of Verapamil HCl at 24 hpi. B, D, F, G, H, I, MARC-145 cells were treated with Verapamil HCl (80 µM) at different time points relative to PRRSV SD16 (MOI＝0.1) infection. progeny virus copies was calculated using standard curve. The data shown are representative of three independent experiments. *** p &lt; 0.001: compared with DMSO-treated cells.Raw data for Figure4 derived from Verapamil HCl inhibits PRRSV infection by blocking Ca2+ influx. MARC-145 cells were seeded into 96-well cell plates and pre-incubated with Verapamil HCl (40 µM or 80 µM) for 2 h. The cells were then infected with 1 MOI PRRSV SD16 for 1 h, with KCl (80 µM) and BAPTA (40 µM) serving as controls. Finally, all cells were treated with Fluo-4 dye-loading solution (100 μL) at 24 hpi and incubated at 37°C for 0.5 h. Calcium flux was detected using a multi-functional microplate reader at Ex/Em = 485/526 nm. The data shown are representative of three independent experiments. *** p &lt; 0.001: compared with PRRSV-infected cells.Raw data for Figure5 derived from Verapamil HCl antagonizes PRRSV-induced inflammatory responses in PAMs. PAMs were treated with 40 µM or 80 µM Verapamil HCl in the presence of PRRSV SD16 (MOI = 0.1). The production of IL-6, IL-8, IL1-β, and TNF-α level at 24 hpi was calculated using OD450 nm by standard curve. The data shown are representative of three independent experiments.**p &lt; 0.01 and ***p &lt; 0.001: compared with DMSO+PRRSV-treated cells.Raw data for Figure6 derived from Verapamil HCl upregulates Nrf2 expression to suppress PRRSV replication. (A, B, D, E) PAMs were treated with various concentrations of Verapamil HCl (10 µM, 20 µM, 40 µM, and 80 µM) or cobalt protoporphyrin (CoPP) (40 µM) for 24 h, with or without PRRSV SD16 infection (MOI = 0.1) for 1 h. (G, H) PAMs were transfected with siHO-1/siNrf2 or siNC and then infected with PRRSV SD16 in the presence or absence of Verapamil HCl (80 µM) for 24 h the relative quantification of Nrf2, HO-1, and Keap1 expression was performed using the 2−∆∆Ct method. GAPDH served as the internal control. The data shown are representative of three independent experiments. **p &lt; 0.01, ***p &lt; 0.001, and ns: not significant.Raw data for Figure7 derived from Verapamil HCl exerts anti-PRRSV effects by inducing an IFN-α/β antiviral response. PAMs were pretreated with Verapamil HCl (40 µM or 80 µM) for 2 h and then incubated without or with PRRSV SD16 (MOI = 0.1 MOI). The levels of IFN-α/β in the supernatant was calculated using OD450 nm by standard curve. The data shown are representative of three independent experiments. *p &lt; 0.05 and ***p &lt; 0.001: compared with 0 µM Verapamil HCl-treated cells.Raw data for Figure8 derived from Verapamil HCl activates the p38/Nrf2/HO-1 cell signaling pathway. MARC-145 cells were incubated with a mixture of Verapamil HCl and a JNK inhibitor (SP600125), ERK1/2 inhibitor (PD98059), or p38 inhibitor (SB203580) for 2 h, and these cells were harvested to analyze Nrf2 and HO-1 expression using qPCR by the 2−∆∆Ct method. MARC-145 cells were incubated with a mixture of Verapamil HCl (80 µM) and a p38 inhibitor (SB203580) for 2 h prior to infection with PRRSV SD16 (MOI = 0.1). The Nrf2 and N protein expression in these cells was analyzed at 24 hpi using qPCR by the 2−∆∆Ct method. GAPDH served as the internal control. All the data shown are representative of three independent experiments. ** p &lt; 0.01 and *** p &lt; 0.001.Raw data for Figure9 derived from Verapamil HCl protects against PRRSV replication in piglets. Fifteen 6-week-old piglets (free of PRRSV) were randomly divided into three groups (n=5). Group 1 was exposed to SD16 only, Group 2 was challenged with SD16 in the presence of Verapamil HCl, and Group 3 was treated with DMSO. Group 1 and Group 2 piglets were challenged with PRRSV SD16 (3×105 TCID50) via nasal drip. All pigs had free access to feed and water and were housed in separate rooms to prevent cross-contamination. Group 1 and Group 3 received MEM containing 0.1% DMSO (5 mL orally) and Group 2 received 20 mg/kg Verapamil HCl diluted in 5 mL of MEM orally on -1, 1, 3, 5 day-post challenge (DPC). Serum samples were collected on days 3, 7, 10, 14, and 21day after the virus challenge, and the copies of SD16 were calculated by standard curve using qPCR, also the levels of IFN-α, IFN-β production level were measured by standard curve using ELISA kits. *p &lt; 0.05, **p &lt; 0.01, and ***p &lt; 0.001.  
description: Raw data for Figure1B derived from Cytotoxicity of Verapamil HCl against MARC-145 cells, PK-15CD163 cells, and PAMs, these cells were treated with different doses of Verapamil HCl (10, 20, 40, 60, 80, and 100 µM) for 24 h, cell viability was detected using the CCK-8 kit. The viable cell count was assessed through absorbance measurements at 450 nm using a microplate reader. In relative cell viability calculations, the untreated cells served as blank controls.Raw data for Figure1D, F, H derived from Verapamil HCl suppresses PRRSV SD16 infection in a dose-dependent manner. MARC-145 cells, PK-15CD163 cells, and PAMs were infected with PRRSV SD16 (MOI = 0.1) pre-incubated with Verapamil HCl (5, 10, 20, 40, and 80 µM). progeny virus copies was calculated using standard curve. The data shown are representative of three independent experiments and were analyzed using Student’s t-tests. ** p &lt; 0.01 and *** p &lt; 0.001: compared with 0 µM Verapamil HCl-treated cells. Raw data for Figure2 derived from anti-PRRSV effects of Verapamil HCl. MARC-145 cells and PAMs were incubated with 0µM or 80 µM Verapamil HCl for 2 h. The cells were infected with various PRRSV-1 and PRRSV-2 strains at 0.1 MOI for 1 h at 37°C. At 24 hpi, the cells were harvested to measure PRRSV N protein expression at the transcription and protein levels using qPCR, GAPDH served as the reference gene, the relative quantification of ORF-7 expression was performed using the 2−∆∆Ct method. Raw data for Figure3 derived from the stage of Verapamil HCl action during the PRRSV life cycle. A, PRRSV SD16 was pre-incubated with Verapamil HCl (80 µM) or DMSO for 2 h at 37°C. Then, this mixture was inoculated into MARC-145 cells. progeny virus copies was calculated using standard curve to evaluate the virucidal activity of Verapamil HCl at 24 hpi. B, D, F, G, H, I, MARC-145 cells were treated with Verapamil HCl (80 µM) at different time points relative to PRRSV SD16 (MOI＝0.1) infection. progeny virus copies was calculated using standard curve. The data shown are representative of three independent experiments. *** p &lt; 0.001: compared with DMSO-treated cells.Raw data for Figure4 derived from Verapamil HCl inhibits PRRSV infection by blocking Ca2+ influx. MARC-145 cells were seeded into 96-well cell plates and pre-incubated with Verapamil HCl (40 µM or 80 µM) for 2 h. The cells were then infected with 1 MOI PRRSV SD16 for 1 h, with KCl (80 µM) and BAPTA (40 µM) serving as controls. Finally, all cells were treated with Fluo-4 dye-loading solution (100 μL) at 24 hpi and incubated at 37°C for 0.5 h. Calcium flux was detected using a multi-functional microplate reader at Ex/Em = 485/526 nm. The data shown are representative of three independent experiments. *** p &lt; 0.001: compared with PRRSV-infected cells.Raw data for Figure5 derived from Verapamil HCl antagonizes PRRSV-induced inflammatory responses in PAMs. PAMs were treated with 40 µM or 80 µM Verapamil HCl in the presence of PRRSV SD16 (MOI = 0.1). The production of IL-6, IL-8, IL1-β, and TNF-α level at 24 hpi was calculated using OD450 nm by standard curve. The data shown are representative of three independent experiments.**p &lt; 0.01 and ***p &lt; 0.001: compared with DMSO+PRRSV-treated cells.Raw data for Figure6 derived from Verapamil HCl upregulates Nrf2 expression to suppress PRRSV replication. (A, B, D, E) PAMs were treated with various concentrations of Verapamil HCl (10 µM, 20 µM, 40 µM, and 80 µM) or cobalt protoporphyrin (CoPP) (40 µM) for 24 h, with or without PRRSV SD16 infection (MOI = 0.1) for 1 h. (G, H) PAMs were transfected with siHO-1/siNrf2 or siNC and then infected with PRRSV SD16 in the presence or absence of Verapamil HCl (80 µM) for 24 h the relative quantification of Nrf2, HO-1, and Keap1 expression was performed using the 2−∆∆Ct method. GAPDH served as the internal control. The data shown are representative of three independent experiments. **p &lt; 0.01, ***p &lt; 0.001, and ns: not significant.Raw data for Figure7 derived from Verapamil HCl exerts anti-PRRSV effects by inducing an IFN-α/β antiviral response. PAMs were pretreated with Verapamil HCl (40 µM or 80 µM) for 2 h and then incubated without or with PRRSV SD16 (MOI = 0.1 MOI). The levels of IFN-α/β in the supernatant was calculated using OD450 nm by standard curve. The data shown are representative of three independent experiments. *p &lt; 0.05 and ***p &lt; 0.001: compared with 0 µM Verapamil HCl-treated cells.Raw data for Figure8 derived from Verapamil HCl activates the p38/Nrf2/HO-1 cell signaling pathway. MARC-145 cells were incubated with a mixture of Verapamil HCl and a JNK inhibitor (SP600125), ERK1/2 inhibitor (PD98059), or p38 inhibitor (SB203580) for 2 h, and these cells were harvested to analyze Nrf2 and HO-1 expression using qPCR by the 2−∆∆Ct method. MARC-145 cells were incubated with a mixture of Verapamil HCl (80 µM) and a p38 inhibitor (SB203580) for 2 h prior to infection with PRRSV SD16 (MOI = 0.1). The Nrf2 and N protein expression in these cells was analyzed at 24 hpi using qPCR by the 2−∆∆Ct method. GAPDH served as the internal control. All the data shown are representative of three independent experiments. ** p &lt; 0.01 and *** p &lt; 0.001.Raw data for Figure9 derived from Verapamil HCl protects against PRRSV replication in piglets. Fifteen 6-week-old piglets (free of PRRSV) were randomly divided into three groups (n=5). Group 1 was exposed to SD16 only, Group 2 was challenged with SD16 in the presence of Verapamil HCl, and Group 3 was treated with DMSO. Group 1 and Group 2 piglets were challenged with PRRSV SD16 (3×105 TCID50) via nasal drip. All pigs had free access to feed and water and were housed in separate rooms to prevent cross-contamination. Group 1 and Group 3 received MEM containing 0.1% DMSO (5 mL orally) and Group 2 received 20 mg/kg Verapamil HCl diluted in 5 mL of MEM orally on -1, 1, 3, 5 day-post challenge (DPC). Serum samples were collected on days 3, 7, 10, 14, and 21day after the virus challenge, and the copies of SD16 were calculated by standard curve using qPCR, also the levels of IFN-α, IFN-β production level were measured by standard curve using ELISA kits. *p &lt; 0.05, **p &lt; 0.01, and ***p &lt; 0.001.