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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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qrt pcr  (Bio-Rad)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hisxript ii  (Vazyme Biotech Co)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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quantitative reverse transcriptase polymerase chain reaction qrt pcr rna  (Qiagen)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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qrt pcr analysis  (Bio-Rad)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
Qrt Pcr Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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primescript qrt pcr kit  (TaKaRa)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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qrt pcr analyses  (Bio-Rad)
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction <t>(qRT-PCR)</t> in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.
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in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction (qRT-PCR) in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.

Journal: Memórias do Instituto Oswaldo Cruz

Article Title: Enhancement of simeprevir and sofosbuvir anti-ZIKV activity by iNOS-derived NO in vitro

doi: 10.1590/0074-02760250315

Figure Lengend Snippet: in vitro evaluation of the protein expression of simeprevir and sofosbuvir in Zika virus (ZIKV)-infected cells. Quantitative content of inducible nitric oxide synthase (iNOS) (A), NF-κB (B), and ARG-I (C) mRNA by quantitative real-time polymerase chain reaction (qRT-PCR) in Vero CCL-81 cells infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), and dimethyl sulfoxide (DMSO) (0.05%). The data are shown as the mean ± SDM of three independent experiments (n = 3). P values of 0.05 or less were considered significant, as indicated by the asterisks. ns indicates not significant. (D) Vero CCL-81 cells were infected with ZIKV (2x10 7 RNA copies/µL) and/or treated with L-NAME and L-norvaline (10 mmol/L), simeprevir and sofosbuvir (10 µmol/L), or DMSO (0.05%). Cell supernatants were collected to measure nitrite production through the Griess method. The data are shown as the mean ± SDM of three independent experiments (n = 9). P values of 0.05 or less were considered significant, as indicated by the asterisks.

Article Snippet: A one-step qRT-PCR kit, RNAdirect Real-time PCR Master Mix (TOYOBO, Japan), and a TaqMan probe were used to quantify the viral RNA produced.

Techniques: In Vitro Evaluation, Expressing, Virus, Infection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR