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coronavirus  (Novus Biologicals)


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    Novus Biologicals coronavirus
    Patient Demographics
    Coronavirus, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/fipv3+70/Coronavirus+Antibody+(FIPV3-70)+-+BSA+Free/pmc07102671-58-16-18
    Average 91 stars, based on 6 article reviews
    coronavirus - by Bioz Stars, 2026-09
    91/100 stars

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    1) Product Images from "Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection"

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection

    Journal: The Journal of Heart and Lung Transplantation

    doi: 10.1016/j.healun.2019.12.009

    Patient Demographics
    Figure Legend Snippet: Patient Demographics

    Techniques Used: Infection

    Lung-associated SAgs and viral antigens were demonstrable in exosomes isolated from patients with RVI. Exosomes isolated from serum samples of patients with RVI and from stable LTxRs were used to detect the presence of lung-associated SAgs and viral antigens using immunoblot. The results showed a significant increase in lung-associated antigens and viral antigens. ( A ) RSV ( n = 10), coronavirus ( n = 12), and rhinovirus ( n = 12) in respective patients with viral infection compared with stable LTxRs ( n = 30). ( B ) Graphical representation shows the optical density of lung-associated SAgs and viral antigens measured in RVI and stable LTxRs using ImageJ software. Optical density of lung SAgs and viral antigens were normalized with exosomes specific marker CD9. The presence of lung SAgs and viral antigens in the exosomes was compared between the cohorts using Mann–Whitney test. Asterisk indicates statistically significant. Col-V, collagen-V; CV, coronavirus; Kα1T, K-alpha-1 tubulin; LTxR, lung transplant recipient; RSV, respiratory syncytial virus; RV, rhinovirus; RVI, respiratory viral infection; SAg, self-antigen.
    Figure Legend Snippet: Lung-associated SAgs and viral antigens were demonstrable in exosomes isolated from patients with RVI. Exosomes isolated from serum samples of patients with RVI and from stable LTxRs were used to detect the presence of lung-associated SAgs and viral antigens using immunoblot. The results showed a significant increase in lung-associated antigens and viral antigens. ( A ) RSV ( n = 10), coronavirus ( n = 12), and rhinovirus ( n = 12) in respective patients with viral infection compared with stable LTxRs ( n = 30). ( B ) Graphical representation shows the optical density of lung-associated SAgs and viral antigens measured in RVI and stable LTxRs using ImageJ software. Optical density of lung SAgs and viral antigens were normalized with exosomes specific marker CD9. The presence of lung SAgs and viral antigens in the exosomes was compared between the cohorts using Mann–Whitney test. Asterisk indicates statistically significant. Col-V, collagen-V; CV, coronavirus; Kα1T, K-alpha-1 tubulin; LTxR, lung transplant recipient; RSV, respiratory syncytial virus; RV, rhinovirus; RVI, respiratory viral infection; SAg, self-antigen.

    Techniques Used: Isolation, Western Blot, Infection, Software, Marker, MANN-WHITNEY, Virus

    Related Articles

    Infection:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Isolation:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Western Blot:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Software:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Marker:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    MANN-WHITNEY:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Virus:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).

    Polymerase Chain Reaction:

    Article Title: Exploiting the Achilles' Heel of Viral RNA Processing to Develop Novel Antivirals.
    Article Snippet: espectively, as previously detailed [17]. To measure the effect of the compound on HCoV-229E, HCoV-OC43, or SARS-CoV-2 nucleocapsid (N) protein expression, mouse α-coronavirus antibody clone FIPV3-70 (Novus Biologicals, Cat: NB10064754), mouse α-coronavirus group antigen antibody clone 542-7D (Millipore Sigma, Cat: MAB9013), or rabbit anti-SARS-CoV-2 nucleocapsid antibody (SinoBiological, Cat: 4014

    Article Title: Respiratory viral infection in lung transplantation induces exosomes that trigger chronic rejection
    Article Snippet: The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).The membrane was blocked with 5% non-fat milk in 1x phosphate buffered saline and was probed with exosome-specific marker CD9 (312102, BioLegend), Col-V (ab7046, Abcam, Cambridge, United Kingdom), and Kα1T (sc-12462-R, Santa Cruz Biotechnology, Dallas, TX).. 20S proteasome subunit α3 (sc-58414, Santa Cruz Biotechnology), rhinovirus VP3 (MA5-18249, Thermo Fisher Scientific, Waltham, MA), coronavirus (NB100-64754, Novus Biologicals, Littleton, CO), and RSV glycoprotein G (7950-0980, Bio-Rad Laboratories, Hercules, CA) were used as primary Abs; secondary Abs conjugated with horseradish peroxidase (HRP) were used specific to primary Ab.. The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).The blots were washed with PBS Tween (Thermo Fisher Scientific), developed using chemiluminescent HRP substrate (WBKLS0500, MilliporeSigma, Burlington, MA), and exposed using Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).



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    Thermo Fisher bcov-nucleocapsid mouse anti-bovine monoclonal (clone: fipv3-70; cat. no. ma1-82189)
    The overexpression of the bovine miRNA16a inhibited <t>BCoV</t> replication on the viral genome copy numbers and the viral infectivity levels. ( A ) In silico prediction of miRNA16atargeting the BCoV spike gene at two different sites. The folding energy represents the binding energy of the miRNA with the target region. ( B ) Multiple sequence alignment shows that the miRNA16a/Spike binding region (indicated in red box) is conserved among nine different BCoV/Ent and BCoV/Resp isolates. ( C ) qRT-PCR analysis demonstrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected MDBK cells. ( D ) qRT-PCR analysis illustrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected BEC cells. ( E ) Viral plaque assay indicating the infectivity level of the BCoV enteric isolate in scrambled- and miRNA16a-transfected MDBK cells. ( F ) Western blot analysis of <t>BCoV-nucleocapsid</t> (BCoV-N) and BCoV-spike (BCoV-S) in the MDBK cells transfected with scrambled or miRNA16a. ( G ) Western blot band density of the BCoV-N protein normalized to that of β-actin in the MDBK cells. ( H ) Western blot band density of the BCoV-S protein normalized to that of β-actin in the MDBK cells. ( I ) BEC cells were transfected with miRNA-Scr and miRNA16a, and western blot analysis was used to assess the protein expression of BCoV-N and BCoV-S. ( J ) Western blot band density of BCoV-N protein normalized to that of β-actin in BEC cells. ( K ) Western blot band density of BCoV-S protein normalized to that of β-actin in BEC cells. All the experiments were performed in triplicate. The significance of the data was determined by one-way ANOVA with Dunnett’s multiple comparison test and indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
    Bcov Nucleocapsid Mouse Anti Bovine Monoclonal (Clone: Fipv3 70; Cat. No. Ma1 82189), supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bcov-nucleocapsid mouse anti-bovine monoclonal (clone: fipv3-70; cat. no. ma1-82189) - by Bioz Stars, 2026-09
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    The overexpression of the bovine miRNA16a inhibited BCoV replication on the viral genome copy numbers and the viral infectivity levels. ( A ) In silico prediction of miRNA16atargeting the BCoV spike gene at two different sites. The folding energy represents the binding energy of the miRNA with the target region. ( B ) Multiple sequence alignment shows that the miRNA16a/Spike binding region (indicated in red box) is conserved among nine different BCoV/Ent and BCoV/Resp isolates. ( C ) qRT-PCR analysis demonstrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected MDBK cells. ( D ) qRT-PCR analysis illustrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected BEC cells. ( E ) Viral plaque assay indicating the infectivity level of the BCoV enteric isolate in scrambled- and miRNA16a-transfected MDBK cells. ( F ) Western blot analysis of BCoV-nucleocapsid (BCoV-N) and BCoV-spike (BCoV-S) in the MDBK cells transfected with scrambled or miRNA16a. ( G ) Western blot band density of the BCoV-N protein normalized to that of β-actin in the MDBK cells. ( H ) Western blot band density of the BCoV-S protein normalized to that of β-actin in the MDBK cells. ( I ) BEC cells were transfected with miRNA-Scr and miRNA16a, and western blot analysis was used to assess the protein expression of BCoV-N and BCoV-S. ( J ) Western blot band density of BCoV-N protein normalized to that of β-actin in BEC cells. ( K ) Western blot band density of BCoV-S protein normalized to that of β-actin in BEC cells. All the experiments were performed in triplicate. The significance of the data was determined by one-way ANOVA with Dunnett’s multiple comparison test and indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Journal: Scientific Reports

    Article Title: The dual actions of miRNA16a in restricting Bovine Coronavirus replication through downregulation of Furin and enhancing the host immune response

    doi: 10.1038/s41598-024-80708-4

    Figure Lengend Snippet: The overexpression of the bovine miRNA16a inhibited BCoV replication on the viral genome copy numbers and the viral infectivity levels. ( A ) In silico prediction of miRNA16atargeting the BCoV spike gene at two different sites. The folding energy represents the binding energy of the miRNA with the target region. ( B ) Multiple sequence alignment shows that the miRNA16a/Spike binding region (indicated in red box) is conserved among nine different BCoV/Ent and BCoV/Resp isolates. ( C ) qRT-PCR analysis demonstrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected MDBK cells. ( D ) qRT-PCR analysis illustrating the genome viral load of BCoV in scrambled- and miRNA16a-transfected BEC cells. ( E ) Viral plaque assay indicating the infectivity level of the BCoV enteric isolate in scrambled- and miRNA16a-transfected MDBK cells. ( F ) Western blot analysis of BCoV-nucleocapsid (BCoV-N) and BCoV-spike (BCoV-S) in the MDBK cells transfected with scrambled or miRNA16a. ( G ) Western blot band density of the BCoV-N protein normalized to that of β-actin in the MDBK cells. ( H ) Western blot band density of the BCoV-S protein normalized to that of β-actin in the MDBK cells. ( I ) BEC cells were transfected with miRNA-Scr and miRNA16a, and western blot analysis was used to assess the protein expression of BCoV-N and BCoV-S. ( J ) Western blot band density of BCoV-N protein normalized to that of β-actin in BEC cells. ( K ) Western blot band density of BCoV-S protein normalized to that of β-actin in BEC cells. All the experiments were performed in triplicate. The significance of the data was determined by one-way ANOVA with Dunnett’s multiple comparison test and indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

    Article Snippet: The primary antibodies were used to detect the expression levels of the BCoV-nucleocapsid mouse anti-bovine monoclonal (clone: FIPV3-70; Cat. No. MA1-82,189), BCoV-spike rabbit anti-bovine polyclonal (cat. no. PA5-117562), and β-actin rabbit anti-bovine polyclonal (Catalogue number: PA1-46296) antibodies were purchased from Invitrogen.

    Techniques: Over Expression, Infection, In Silico, Binding Assay, Sequencing, Quantitative RT-PCR, Transfection, Viral Plaque Assay, Western Blot, Expressing, Comparison