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wnt inhibitor e7449  (Charles River Laboratories)

 
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    Charles River Laboratories wnt inhibitor e7449
    A Calu-3 cells were pre-treated with the Wnt <t>inhibitors</t> for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.
    Wnt Inhibitor E7449, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wnt inhibitor e7449/product/Charles River Laboratories
    Average 90 stars, based on 1 article reviews
    wnt inhibitor e7449 - by Bioz Stars, 2026-02
    90/100 stars

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    1) Product Images from "The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses"

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    Journal: npj Viruses

    doi: 10.1038/s44298-024-00018-4

    A Calu-3 cells were pre-treated with the Wnt inhibitors for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.
    Figure Legend Snippet: A Calu-3 cells were pre-treated with the Wnt inhibitors for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.

    Techniques Used: Infection, Concentration Assay, Plaque Assay, Comparison, Quantitative RT-PCR, Western Blot, Virus

    Calu-3 cells were pre-treated with the indicated concentrations of Wnt inhibitors IWP-O1, KYA1797K and Pyrvinium for 24-h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, cell media were collected and subjected to plaque assay to determine viral titers. Viabilities of cells treated with Wnt inhibitors for 48 h in the absence of infection were determined by cytotoxicity assay. A Relative average viral titers obtained from three independent experiments are shown as are the relative cell viabilities of cells treated with Wnt inhibitor for 48 h in the absence of infection. EC 50 and CC 50 values were determined and then used to calculate the selectivity indexes (CC 50 /EC 50 ) for each drug. ( B – G ) Calu-3 cells were pre-treated with IWP-O1 (10 nM, 100 nM and 1 μM), KYA1797K (10 nM, 100 nM, 1 μM and 10 μM) and Pyrvinium (1 nM, 10 nM and 100 nM) for 24 h and then infected with SARS-CoV-2 variants ( B D614G, C UK B.1.1.7 (Alpha), D SA B.1.351 (Beta), E Brazil P.1 (Gamma), F India B.1.617.2 (Delta) and G B.1.1.529 (Omicron)) using an MOI of 0.5. Twenty-four hours later, cell media were subjected to plaque assay. Viral titers from three independent experiments were determined and averaged. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Figure Legend Snippet: Calu-3 cells were pre-treated with the indicated concentrations of Wnt inhibitors IWP-O1, KYA1797K and Pyrvinium for 24-h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, cell media were collected and subjected to plaque assay to determine viral titers. Viabilities of cells treated with Wnt inhibitors for 48 h in the absence of infection were determined by cytotoxicity assay. A Relative average viral titers obtained from three independent experiments are shown as are the relative cell viabilities of cells treated with Wnt inhibitor for 48 h in the absence of infection. EC 50 and CC 50 values were determined and then used to calculate the selectivity indexes (CC 50 /EC 50 ) for each drug. ( B – G ) Calu-3 cells were pre-treated with IWP-O1 (10 nM, 100 nM and 1 μM), KYA1797K (10 nM, 100 nM, 1 μM and 10 μM) and Pyrvinium (1 nM, 10 nM and 100 nM) for 24 h and then infected with SARS-CoV-2 variants ( B D614G, C UK B.1.1.7 (Alpha), D SA B.1.351 (Beta), E Brazil P.1 (Gamma), F India B.1.617.2 (Delta) and G B.1.1.529 (Omicron)) using an MOI of 0.5. Twenty-four hours later, cell media were subjected to plaque assay. Viral titers from three independent experiments were determined and averaged. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Techniques Used: Infection, Plaque Assay, Cytotoxicity Assay, Comparison

    A A549 cells were treated with DMSO alone or ten different commercially available drugs/compounds (1 μM) that block Wnt/β-catenin signaling. Cells were fixed at 24- and 48-h post-drug treatment and processed for confocal microscopy using an antibody against PEX14 to label peroxisomes and CellMask TM to label the plasma membrane. The numbers of peroxisomes in cells were determined using Volocity software. The peroxisome density (number/μm 3 ) was calculated by dividing the number of peroxisomes by the estimated cell volume. For each sample, peroxisome densities were determined using a minimum of 20 cells. Data from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; ** P < 0.01. B Calu-3 cells were treated with the indicated Wnt inhibitors (1 μM) or Pyrvinium (100 nM) for 48-h and then total RNA, including small RNAs, were extracted from the samples, and relative levels of miRNAs were determined by RT-qPCR. The average relative levels of miRNAs (normalized to snRNU6) from three independent experiments were determined. Two-way ANOVA with Bonferroni post-hoc tests was used to determine significance between samples treated with DMSO and Wnt inhibitors. Error bars represent standard errors of the mean. *** P < 0.001.
    Figure Legend Snippet: A A549 cells were treated with DMSO alone or ten different commercially available drugs/compounds (1 μM) that block Wnt/β-catenin signaling. Cells were fixed at 24- and 48-h post-drug treatment and processed for confocal microscopy using an antibody against PEX14 to label peroxisomes and CellMask TM to label the plasma membrane. The numbers of peroxisomes in cells were determined using Volocity software. The peroxisome density (number/μm 3 ) was calculated by dividing the number of peroxisomes by the estimated cell volume. For each sample, peroxisome densities were determined using a minimum of 20 cells. Data from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; ** P < 0.01. B Calu-3 cells were treated with the indicated Wnt inhibitors (1 μM) or Pyrvinium (100 nM) for 48-h and then total RNA, including small RNAs, were extracted from the samples, and relative levels of miRNAs were determined by RT-qPCR. The average relative levels of miRNAs (normalized to snRNU6) from three independent experiments were determined. Two-way ANOVA with Bonferroni post-hoc tests was used to determine significance between samples treated with DMSO and Wnt inhibitors. Error bars represent standard errors of the mean. *** P < 0.001.

    Techniques Used: Blocking Assay, Confocal Microscopy, Clinical Proteomics, Membrane, Software, Quantitative RT-PCR

    A549 cells were treated with DMSO alone or 10 different commercially available drugs (1 μM) that block Wnt/β-catenin signaling. Twenty-four hours later, cells were infected with 100 HAU/ml of Sendai virus after which total cellular RNA was harvested 8- or 16-h post infection (hpi) and then subjected to qRT-PCR to determine relative levels mRNA encoding ( A ) type I (IFNβ) and ( B ) type III (IFNλ2) interferons. Values from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Figure Legend Snippet: A549 cells were treated with DMSO alone or 10 different commercially available drugs (1 μM) that block Wnt/β-catenin signaling. Twenty-four hours later, cells were infected with 100 HAU/ml of Sendai virus after which total cellular RNA was harvested 8- or 16-h post infection (hpi) and then subjected to qRT-PCR to determine relative levels mRNA encoding ( A ) type I (IFNβ) and ( B ) type III (IFNλ2) interferons. Values from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Techniques Used: Blocking Assay, Infection, Virus, Quantitative RT-PCR

    A Wnt/β-catenin inhibitors were administered intranasally to female Balb/c mice once daily from day-2 to day +2 relative to SARS-CoV-2 infection. Lung tissues were collected on 4 days post infection (DPI) and processed for virus titration, viral RNA load measurement and histopathology analysis. Mice health and weight were monitored daily for signs of infection and morbidity for up to 14 days. B Virus titers (PFU per g of tissue) from lungs of infected mice at 4 DPI ( n = 10 mice per group) were determined by plaque assay. Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Viral RNA load (RNA copies (N2) per μg of total RNA) from lungs of infected mice at 4 DPI ( n = 10 mice per group). Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. D Weight changes in infected mice treated with DMSO control or Wnt inhibitors KYA1797K and E7449 over the course of 14 days. E Histopathological analysis of mouse lung tissues. Uninfected (i, ii), infected, DMSO treated (iii, iv), infected, KYA1797K treated (v, vi) and infected, E7449 treated (vii, viii). Panels ii, iv, vi and viii are enlargements of boxed areas in panels i, iii, v, and vii respectively. Scale bars, 50 μm. SFM serum-free media.
    Figure Legend Snippet: A Wnt/β-catenin inhibitors were administered intranasally to female Balb/c mice once daily from day-2 to day +2 relative to SARS-CoV-2 infection. Lung tissues were collected on 4 days post infection (DPI) and processed for virus titration, viral RNA load measurement and histopathology analysis. Mice health and weight were monitored daily for signs of infection and morbidity for up to 14 days. B Virus titers (PFU per g of tissue) from lungs of infected mice at 4 DPI ( n = 10 mice per group) were determined by plaque assay. Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Viral RNA load (RNA copies (N2) per μg of total RNA) from lungs of infected mice at 4 DPI ( n = 10 mice per group). Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. D Weight changes in infected mice treated with DMSO control or Wnt inhibitors KYA1797K and E7449 over the course of 14 days. E Histopathological analysis of mouse lung tissues. Uninfected (i, ii), infected, DMSO treated (iii, iv), infected, KYA1797K treated (v, vi) and infected, E7449 treated (vii, viii). Panels ii, iv, vi and viii are enlargements of boxed areas in panels i, iii, v, and vii respectively. Scale bars, 50 μm. SFM serum-free media.

    Techniques Used: Infection, Virus, Titration, Histopathology, Plaque Assay, Comparison, Control

    A549 cells were treated with Wnt/β-catenin inhibitors (IWP-O1 (1 μM), KYA1797K (1 μM), Pyrvinium (100 nM), E7449 (1 μM)) or DMSO alone for 24 h, after which the cells were infected with 0.1 MOI of Zika virus (ZIKV) for 48-h or Mayaro virus (MAYV) for 24-h. Virus-containing media were subjected to plaque assays and total RNA extracted from cells was subjected to qRT-PCR to determine relative levels of viral RNA. Average viral titers ( A , C ) and genomic RNA levels ( B , D ) from drug-treated cells from 3 independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; *** P < 0.001.
    Figure Legend Snippet: A549 cells were treated with Wnt/β-catenin inhibitors (IWP-O1 (1 μM), KYA1797K (1 μM), Pyrvinium (100 nM), E7449 (1 μM)) or DMSO alone for 24 h, after which the cells were infected with 0.1 MOI of Zika virus (ZIKV) for 48-h or Mayaro virus (MAYV) for 24-h. Virus-containing media were subjected to plaque assays and total RNA extracted from cells was subjected to qRT-PCR to determine relative levels of viral RNA. Average viral titers ( A , C ) and genomic RNA levels ( B , D ) from drug-treated cells from 3 independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; *** P < 0.001.

    Techniques Used: Infection, Virus, Quantitative RT-PCR, Comparison



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    wnt inhibitor e7449  (Charles River Laboratories)
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    Charles River Laboratories wnt inhibitor e7449
    A Calu-3 cells were pre-treated with the Wnt <t>inhibitors</t> for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.
    Wnt Inhibitor E7449, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wnt inhibitor e7449/product/Charles River Laboratories
    Average 90 stars, based on 1 article reviews
    wnt inhibitor e7449 - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

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    A Calu-3 cells were pre-treated with the Wnt inhibitors for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: A Calu-3 cells were pre-treated with the Wnt inhibitors for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using MOI of 0.5. Apart from Pyrvinium (100 nM), the final concentration of all other Wnt inhibitors was 1 μM. Twenty-four hours later, media were collected and subjected to plaque assay to determine viral titers. Data shown are averaged from three independent experiments. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. B Total RNA extracted from infected cells at 24 h post-infection (hpi) was subjected to qRT-PCR analysis to determine relative (to actin mRNA) levels of SARS-CoV-2 viral RNA. Data averaged from three independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Cell lysates harvested at 24 hpi were processed for immunoblot analyses with antibodies to SARS-CoV-2 spike protein and actin. D Primary normal human bronchial epithelial (NHBE) cells were pre-treated with Wnt inhibitors (IWP-O1, NCB-0846, Pyrvinium and SM04690 used at 100 nM, others used at 1 μM) for 24 h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, virus-containing media were collected and then subjected to plaque assay to determine viral titers. The average titers from three independent experiments are shown. Dashed line represents limit of detection by plaque assay. Error bars represent the standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Infection, Concentration Assay, Plaque Assay, Comparison, Quantitative RT-PCR, Western Blot, Virus

    Calu-3 cells were pre-treated with the indicated concentrations of Wnt inhibitors IWP-O1, KYA1797K and Pyrvinium for 24-h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, cell media were collected and subjected to plaque assay to determine viral titers. Viabilities of cells treated with Wnt inhibitors for 48 h in the absence of infection were determined by cytotoxicity assay. A Relative average viral titers obtained from three independent experiments are shown as are the relative cell viabilities of cells treated with Wnt inhibitor for 48 h in the absence of infection. EC 50 and CC 50 values were determined and then used to calculate the selectivity indexes (CC 50 /EC 50 ) for each drug. ( B – G ) Calu-3 cells were pre-treated with IWP-O1 (10 nM, 100 nM and 1 μM), KYA1797K (10 nM, 100 nM, 1 μM and 10 μM) and Pyrvinium (1 nM, 10 nM and 100 nM) for 24 h and then infected with SARS-CoV-2 variants ( B D614G, C UK B.1.1.7 (Alpha), D SA B.1.351 (Beta), E Brazil P.1 (Gamma), F India B.1.617.2 (Delta) and G B.1.1.529 (Omicron)) using an MOI of 0.5. Twenty-four hours later, cell media were subjected to plaque assay. Viral titers from three independent experiments were determined and averaged. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: Calu-3 cells were pre-treated with the indicated concentrations of Wnt inhibitors IWP-O1, KYA1797K and Pyrvinium for 24-h and then infected with SARS-CoV-2 (CANADA/VIDO01/2020 strain) using an MOI of 0.5. Twenty-four hours later, cell media were collected and subjected to plaque assay to determine viral titers. Viabilities of cells treated with Wnt inhibitors for 48 h in the absence of infection were determined by cytotoxicity assay. A Relative average viral titers obtained from three independent experiments are shown as are the relative cell viabilities of cells treated with Wnt inhibitor for 48 h in the absence of infection. EC 50 and CC 50 values were determined and then used to calculate the selectivity indexes (CC 50 /EC 50 ) for each drug. ( B – G ) Calu-3 cells were pre-treated with IWP-O1 (10 nM, 100 nM and 1 μM), KYA1797K (10 nM, 100 nM, 1 μM and 10 μM) and Pyrvinium (1 nM, 10 nM and 100 nM) for 24 h and then infected with SARS-CoV-2 variants ( B D614G, C UK B.1.1.7 (Alpha), D SA B.1.351 (Beta), E Brazil P.1 (Gamma), F India B.1.617.2 (Delta) and G B.1.1.529 (Omicron)) using an MOI of 0.5. Twenty-four hours later, cell media were subjected to plaque assay. Viral titers from three independent experiments were determined and averaged. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Infection, Plaque Assay, Cytotoxicity Assay, Comparison

    A A549 cells were treated with DMSO alone or ten different commercially available drugs/compounds (1 μM) that block Wnt/β-catenin signaling. Cells were fixed at 24- and 48-h post-drug treatment and processed for confocal microscopy using an antibody against PEX14 to label peroxisomes and CellMask TM to label the plasma membrane. The numbers of peroxisomes in cells were determined using Volocity software. The peroxisome density (number/μm 3 ) was calculated by dividing the number of peroxisomes by the estimated cell volume. For each sample, peroxisome densities were determined using a minimum of 20 cells. Data from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; ** P < 0.01. B Calu-3 cells were treated with the indicated Wnt inhibitors (1 μM) or Pyrvinium (100 nM) for 48-h and then total RNA, including small RNAs, were extracted from the samples, and relative levels of miRNAs were determined by RT-qPCR. The average relative levels of miRNAs (normalized to snRNU6) from three independent experiments were determined. Two-way ANOVA with Bonferroni post-hoc tests was used to determine significance between samples treated with DMSO and Wnt inhibitors. Error bars represent standard errors of the mean. *** P < 0.001.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: A A549 cells were treated with DMSO alone or ten different commercially available drugs/compounds (1 μM) that block Wnt/β-catenin signaling. Cells were fixed at 24- and 48-h post-drug treatment and processed for confocal microscopy using an antibody against PEX14 to label peroxisomes and CellMask TM to label the plasma membrane. The numbers of peroxisomes in cells were determined using Volocity software. The peroxisome density (number/μm 3 ) was calculated by dividing the number of peroxisomes by the estimated cell volume. For each sample, peroxisome densities were determined using a minimum of 20 cells. Data from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; ** P < 0.01. B Calu-3 cells were treated with the indicated Wnt inhibitors (1 μM) or Pyrvinium (100 nM) for 48-h and then total RNA, including small RNAs, were extracted from the samples, and relative levels of miRNAs were determined by RT-qPCR. The average relative levels of miRNAs (normalized to snRNU6) from three independent experiments were determined. Two-way ANOVA with Bonferroni post-hoc tests was used to determine significance between samples treated with DMSO and Wnt inhibitors. Error bars represent standard errors of the mean. *** P < 0.001.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Blocking Assay, Confocal Microscopy, Clinical Proteomics, Membrane, Software, Quantitative RT-PCR

    A549 cells were treated with DMSO alone or 10 different commercially available drugs (1 μM) that block Wnt/β-catenin signaling. Twenty-four hours later, cells were infected with 100 HAU/ml of Sendai virus after which total cellular RNA was harvested 8- or 16-h post infection (hpi) and then subjected to qRT-PCR to determine relative levels mRNA encoding ( A ) type I (IFNβ) and ( B ) type III (IFNλ2) interferons. Values from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: A549 cells were treated with DMSO alone or 10 different commercially available drugs (1 μM) that block Wnt/β-catenin signaling. Twenty-four hours later, cells were infected with 100 HAU/ml of Sendai virus after which total cellular RNA was harvested 8- or 16-h post infection (hpi) and then subjected to qRT-PCR to determine relative levels mRNA encoding ( A ) type I (IFNβ) and ( B ) type III (IFNλ2) interferons. Values from three independent experiments are shown. Error bars represent standard errors of the mean. Two-way ANOVA with Bonferroni post-hoc tests were used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Blocking Assay, Infection, Virus, Quantitative RT-PCR

    A Wnt/β-catenin inhibitors were administered intranasally to female Balb/c mice once daily from day-2 to day +2 relative to SARS-CoV-2 infection. Lung tissues were collected on 4 days post infection (DPI) and processed for virus titration, viral RNA load measurement and histopathology analysis. Mice health and weight were monitored daily for signs of infection and morbidity for up to 14 days. B Virus titers (PFU per g of tissue) from lungs of infected mice at 4 DPI ( n = 10 mice per group) were determined by plaque assay. Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Viral RNA load (RNA copies (N2) per μg of total RNA) from lungs of infected mice at 4 DPI ( n = 10 mice per group). Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. D Weight changes in infected mice treated with DMSO control or Wnt inhibitors KYA1797K and E7449 over the course of 14 days. E Histopathological analysis of mouse lung tissues. Uninfected (i, ii), infected, DMSO treated (iii, iv), infected, KYA1797K treated (v, vi) and infected, E7449 treated (vii, viii). Panels ii, iv, vi and viii are enlargements of boxed areas in panels i, iii, v, and vii respectively. Scale bars, 50 μm. SFM serum-free media.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: A Wnt/β-catenin inhibitors were administered intranasally to female Balb/c mice once daily from day-2 to day +2 relative to SARS-CoV-2 infection. Lung tissues were collected on 4 days post infection (DPI) and processed for virus titration, viral RNA load measurement and histopathology analysis. Mice health and weight were monitored daily for signs of infection and morbidity for up to 14 days. B Virus titers (PFU per g of tissue) from lungs of infected mice at 4 DPI ( n = 10 mice per group) were determined by plaque assay. Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. C Viral RNA load (RNA copies (N2) per μg of total RNA) from lungs of infected mice at 4 DPI ( n = 10 mice per group). Error bars represent standard errors of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. *** P < 0.001. D Weight changes in infected mice treated with DMSO control or Wnt inhibitors KYA1797K and E7449 over the course of 14 days. E Histopathological analysis of mouse lung tissues. Uninfected (i, ii), infected, DMSO treated (iii, iv), infected, KYA1797K treated (v, vi) and infected, E7449 treated (vii, viii). Panels ii, iv, vi and viii are enlargements of boxed areas in panels i, iii, v, and vii respectively. Scale bars, 50 μm. SFM serum-free media.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Infection, Virus, Titration, Histopathology, Plaque Assay, Comparison, Control

    A549 cells were treated with Wnt/β-catenin inhibitors (IWP-O1 (1 μM), KYA1797K (1 μM), Pyrvinium (100 nM), E7449 (1 μM)) or DMSO alone for 24 h, after which the cells were infected with 0.1 MOI of Zika virus (ZIKV) for 48-h or Mayaro virus (MAYV) for 24-h. Virus-containing media were subjected to plaque assays and total RNA extracted from cells was subjected to qRT-PCR to determine relative levels of viral RNA. Average viral titers ( A , C ) and genomic RNA levels ( B , D ) from drug-treated cells from 3 independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; *** P < 0.001.

    Journal: npj Viruses

    Article Title: The Wnt/β-catenin pathway is important for replication of SARS-CoV-2 and other pathogenic RNA viruses

    doi: 10.1038/s44298-024-00018-4

    Figure Lengend Snippet: A549 cells were treated with Wnt/β-catenin inhibitors (IWP-O1 (1 μM), KYA1797K (1 μM), Pyrvinium (100 nM), E7449 (1 μM)) or DMSO alone for 24 h, after which the cells were infected with 0.1 MOI of Zika virus (ZIKV) for 48-h or Mayaro virus (MAYV) for 24-h. Virus-containing media were subjected to plaque assays and total RNA extracted from cells was subjected to qRT-PCR to determine relative levels of viral RNA. Average viral titers ( A , C ) and genomic RNA levels ( B , D ) from drug-treated cells from 3 independent experiments are shown. Error bars represent standard error of the mean. One-way ANOVA with Dunnett’s multiple comparison test was used to determine significance between samples treated with DMSO and Wnt inhibitors. * P < 0.05; *** P < 0.001.

    Article Snippet: Wnt inhibitors were administered to anesthetized 6–8-week-old female Balb/c mice (Charles River Laboratories Canada) at 3 mg/kg (KYA1797K) or 6 mg/kg (E7449) (in a total volume of 50 μL (25 μL per nare)) on day -2 and -1.

    Techniques: Infection, Virus, Quantitative RT-PCR, Comparison