viral dna Search Results


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  • 95
    Zymo Research viral dna kit
    Viral Dna Kit, supplied by Zymo Research, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher genejet viral dna
    Genejet Viral Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher viral dna
    Viral Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 963 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Advanced Biotechnologies Inc viral dna
    Viral Dna, supplied by Advanced Biotechnologies Inc, used in various techniques. Bioz Stars score: 91/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATUM viral dna
    Viral Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson baculogold viral dna
    Baculogold Viral Dna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pharmingen baculogold viral dna
    Baculogold Viral Dna, supplied by Pharmingen, used in various techniques. Bioz Stars score: 88/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Advanced Biotechnologies Inc p3hr1 viral dna
    P3hr1 Viral Dna, supplied by Advanced Biotechnologies Inc, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    TaKaRa acmnpv viral dna
    Acmnpv Viral Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson bacpak viral dna
    Bacpak Viral Dna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Omega Bio-tek viral dna kit
    Tat-PNA-DR inhibits <t>HBV</t> replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV <t>DNA</t> of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.
    Viral Dna Kit, supplied by Omega Bio-tek, used in various techniques. Bioz Stars score: 88/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abivax viral dna levels
    Tat-PNA-DR inhibits <t>HBV</t> replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV <t>DNA</t> of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.
    Viral Dna Levels, supplied by Abivax, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ATUM m13mp18 viral dna
    Tat-PNA-DR inhibits <t>HBV</t> replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV <t>DNA</t> of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.
    M13mp18 Viral Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare viral dna detection
    Tat-PNA-DR inhibits <t>HBV</t> replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV <t>DNA</t> of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.
    Viral Dna Detection, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Viral Dna, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 2698 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher acmnpv viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Acmnpv Viral Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson bakpak viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Bakpak Viral Dna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression Systems Inc bestbac viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Bestbac Viral Dna, supplied by Expression Systems Inc, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pharmingen nonviable viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Nonviable Viral Dna, supplied by Pharmingen, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pbacpak6 viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
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    Becton Dickinson adeno x viral dna
    <t>DNA</t> damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. <t>RT-qPCR</t> was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P
    Adeno X Viral Dna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Tat-PNA-DR inhibits HBV replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV DNA of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Tat-conjugated Peptide Nucleic Acid Tat-PNA-DR Inhibits Hepatitis B Virus Replication In Vitro and In Vivo by Targeting LTR Direct Repeats of HBV RNA

    doi: 10.1038/mtna.2016.11

    Figure Lengend Snippet: Tat-PNA-DR inhibits HBV replication by binding DR in a sequence-specific manner. ( a , b ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm (mutant of Tat-PNA-DR) against the expression of HBeAg and HBsAg in HepG2.2.15 culture medium. HepG2.2.15 cells were incubated with Tat-PNA-DR or Tat-PNA-DRm for 48 hours. The levels of HBeAg and HBsAg in the culture medium were measured using ELISA. ( c ) Comparison of inhibitory activity on extracellular HBV DNA of Tat-PNA-DR and Tat-PNA-DRm was performed by a real-time PCR. The data above are shown as the mean ± SEM of five independent samples. ( d – g ) Western blot assay was used to measure the effects of Tat-PNA-DR and Tat-PNA-DRm on intracellular HBV proteins. ( h ) Inhibitory activity of Tat-PNA-DR and Tat-PNA-DRm on HBV RNA was measured by northern blot with a DIG-labeled probe. ( i ) In vitro binding assay of Tat-PNA-DR and HBV DR. Native and mutated RNA oligonucleotides corresponding to the HBV DR were synthesized and labeled with biotin. The labeled RNA oligonucleotides were used for in vitro binding assays. Shifted bands indicate the binding between conjugates of Tat-PNA-DR and RNA oligonucleotides.

    Article Snippet: HBV capsid DNA was collected from cytoplasm using a viral DNA kit (OMEGA bio-tek, GA, USA).

    Techniques: Binding Assay, Sequencing, Activity Assay, Mutagenesis, Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot, Northern Blot, Labeling, In Vitro, Synthesized

    Anti-HBV activity of Tat-PNA-DR in HepG2.2.15 cells . ( a , b ) The inhibitory effect of Tat-PNA-DR against HBeAg and HBsAg in HepG2.2.15 culture medium was measured using ELISA. ( c ) The inhibitory effect of Tat-PNA-DR against HBV DNA in HepG2.2.15 culture medium was measured using real-time PCR. Values represent the mean ± SEM of five independent samples. ( d – g ) The inhibitory effect of Tat-PNA-DR against the HBsAg, HBV core, HBx, and HBV RT proteins in HepG2.2.15 cell lysates was determined using western blot analysis. The drug 3TC was used as a control. ( h ) The inhibitory effect of Tat-PNA-DR against HBV DNA in HepG2.2.15 cells was determined using southern blot analysis. The drug 3TC, which is a nucleoside reverse transcriptase inhibitor, was used as a control. ( i ) The inhibitory effect of Tat-PNA-DR against HBV RNA in HepG2.2.15 cells was determined using northern blot analysis. The drug 3TC was used as a control.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Tat-conjugated Peptide Nucleic Acid Tat-PNA-DR Inhibits Hepatitis B Virus Replication In Vitro and In Vivo by Targeting LTR Direct Repeats of HBV RNA

    doi: 10.1038/mtna.2016.11

    Figure Lengend Snippet: Anti-HBV activity of Tat-PNA-DR in HepG2.2.15 cells . ( a , b ) The inhibitory effect of Tat-PNA-DR against HBeAg and HBsAg in HepG2.2.15 culture medium was measured using ELISA. ( c ) The inhibitory effect of Tat-PNA-DR against HBV DNA in HepG2.2.15 culture medium was measured using real-time PCR. Values represent the mean ± SEM of five independent samples. ( d – g ) The inhibitory effect of Tat-PNA-DR against the HBsAg, HBV core, HBx, and HBV RT proteins in HepG2.2.15 cell lysates was determined using western blot analysis. The drug 3TC was used as a control. ( h ) The inhibitory effect of Tat-PNA-DR against HBV DNA in HepG2.2.15 cells was determined using southern blot analysis. The drug 3TC, which is a nucleoside reverse transcriptase inhibitor, was used as a control. ( i ) The inhibitory effect of Tat-PNA-DR against HBV RNA in HepG2.2.15 cells was determined using northern blot analysis. The drug 3TC was used as a control.

    Article Snippet: HBV capsid DNA was collected from cytoplasm using a viral DNA kit (OMEGA bio-tek, GA, USA).

    Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot, Southern Blot, Northern Blot

    DNA damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P

    Journal: bioRxiv

    Article Title: HSATII RNA is induced via E2F3a and a non-canonical ATM-regulated DNA damage response pathway

    doi: 10.1101/2020.05.25.115238

    Figure Lengend Snippet: DNA damage response regulates HSATII expression. (A) ARPE-19 cells were exposed to UV-irradiation (50 J/m 2 ), 100 nM of H 2 O 2 or serum withdrawal. RNA samples were collected at 24 hpt. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. * P

    Article Snippet: DNA was RNase A treated (2 mg/ml; Qiagen) and viral DNA was quantified by qPCR using primers specific for the viral genomic UL44 (Forward: 5’-GTGCGCGCCCGATTTCAATATG-3’, Reverse: 5’-GCTTTCGCGCACAATGTCTTGG-3’) or cellular genomic GAPDH (Forward: 5’-CCCCACACACATGCACTTACC-3’, Reverse: 5’-CCTAGTCCCAGGGCTTTGATT-3’).

    Techniques: Expressing, Irradiation, Quantitative RT-PCR

    DNA damage-induced HSATII RNA enhances motility and proliferation of breast cancer cells. (A) Indicated cell lines, ordered according to the p53 status, were grown to ∼80% confluency and RNA samples were collected. RT-qPCR was performed using specific primers to HSATII RNA. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (B) MCF-7, MCF-10A, MDA-MB-361 and SUM1315MO2 cells were treated with 200 μg/mL of Zeocin. RNA samples were collected at 24 and 96 hpt. RT-qPCR was performed using specific primers to HSATII RNA. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (C) ARPE-1, MCF-7 and SUM315MO2 cells were treated with 200 μg/mL of Zeocin for 24 h. Cells were transferred onto transwell inserts. After 24 hpt, migrated cells were washed, fixed and nuclei stained. The graph presents a number of cells (with their indicated means) migrated through a transwell per FOV from biological replicates. n =9-12. (D) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpi, equal number of cells was transferred onto transwell inserts. 24 h later, migrated cells were washed, fixed and nuclei stained. The graph presents a number of cells (with their indicated means) migrated through a transwell per FOV from biological replicates. n =12. (E) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpi, wounds were created and their closure was monitored at indicated times. Data from biological replicates are presented as a percent of remaining wound width mean ± SD. n=6. (F) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpt, equal number of cells were seeded and cell proliferation was monitored using a colorimetric CellTiter 96 AQueous One Solution Cell Proliferation Assay at the indicated times. Cell proliferation is presented as an increase in a measured absorbance (AU – arbitrary units). n=3. * P

    Journal: bioRxiv

    Article Title: HSATII RNA is induced via E2F3a and a non-canonical ATM-regulated DNA damage response pathway

    doi: 10.1101/2020.05.25.115238

    Figure Lengend Snippet: DNA damage-induced HSATII RNA enhances motility and proliferation of breast cancer cells. (A) Indicated cell lines, ordered according to the p53 status, were grown to ∼80% confluency and RNA samples were collected. RT-qPCR was performed using specific primers to HSATII RNA. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (B) MCF-7, MCF-10A, MDA-MB-361 and SUM1315MO2 cells were treated with 200 μg/mL of Zeocin. RNA samples were collected at 24 and 96 hpt. RT-qPCR was performed using specific primers to HSATII RNA. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (C) ARPE-1, MCF-7 and SUM315MO2 cells were treated with 200 μg/mL of Zeocin for 24 h. Cells were transferred onto transwell inserts. After 24 hpt, migrated cells were washed, fixed and nuclei stained. The graph presents a number of cells (with their indicated means) migrated through a transwell per FOV from biological replicates. n =9-12. (D) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpi, equal number of cells was transferred onto transwell inserts. 24 h later, migrated cells were washed, fixed and nuclei stained. The graph presents a number of cells (with their indicated means) migrated through a transwell per FOV from biological replicates. n =12. (E) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpi, wounds were created and their closure was monitored at indicated times. Data from biological replicates are presented as a percent of remaining wound width mean ± SD. n=6. (F) MDA-MB-231, BT-549 and SUM1315MO2 cells were transfected with NT-LNA or HSATII-LNAs. After 48 hpt, equal number of cells were seeded and cell proliferation was monitored using a colorimetric CellTiter 96 AQueous One Solution Cell Proliferation Assay at the indicated times. Cell proliferation is presented as an increase in a measured absorbance (AU – arbitrary units). n=3. * P

    Article Snippet: DNA was RNase A treated (2 mg/ml; Qiagen) and viral DNA was quantified by qPCR using primers specific for the viral genomic UL44 (Forward: 5’-GTGCGCGCCCGATTTCAATATG-3’, Reverse: 5’-GCTTTCGCGCACAATGTCTTGG-3’) or cellular genomic GAPDH (Forward: 5’-CCCCACACACATGCACTTACC-3’, Reverse: 5’-CCTAGTCCCAGGGCTTTGATT-3’).

    Techniques: Quantitative RT-PCR, Multiple Displacement Amplification, Staining, Transfection, Proliferation Assay

    ATM-based DNA damage response regulates HSATII expression. (A and B) ARPE-19 cells were mock- or HCMV-infected with HCMV (TB40/E-GFP-epi) at 3 TCID50 /cell or treated with 200 μg/mL of zeocin. Cells were stained for γ-H2AX and nuclei were counterstained with the Hoechst stain. Cells were visualized (20X magnification) (A) and % of γ-H2AX-positive cells was calculated (B) using Operetta high-content imaging and analysis system. Scale bars: 100 µm. (C and D) ARPE-19 cells were transfected with ATM, ATR or DNA-PK siRNAs or NT siRNA as a control. After 24 h, cells were infected with TB40/E-GFP-epi at 1 TCID50 per cell (A) or 100 μg/mL of zeocin (B). RNA samples were collected 24 h later. RT-qPCR was performed using specific primers to HSATII, ATM, ATR or DNA-PK transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (E) A-T(+) and A-T(-) cells were infected with TB40/E-GFP-epi at 1 TCID50 per cell, 200 μg/mL of zeocin or 200 μM of etoposide. RNA samples were collected 24 h later. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (F) ARPE-19 cells were treated with DMSO, 20 μM of either Ku-55933 or AZ31 for 2 h before zeocin was added at the concentration of 100 μg/mL. RNA samples were coll ected 24 hpt. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (G) ARPE-19 cells were treated with various concentrations of Ku-55933 or AZ31. At 24 hpt, cell viability was assessed. Data is presented as % viable cells, were averaged from at least three independent experiments and are presented as mean (SD). * P

    Journal: bioRxiv

    Article Title: HSATII RNA is induced via E2F3a and a non-canonical ATM-regulated DNA damage response pathway

    doi: 10.1101/2020.05.25.115238

    Figure Lengend Snippet: ATM-based DNA damage response regulates HSATII expression. (A and B) ARPE-19 cells were mock- or HCMV-infected with HCMV (TB40/E-GFP-epi) at 3 TCID50 /cell or treated with 200 μg/mL of zeocin. Cells were stained for γ-H2AX and nuclei were counterstained with the Hoechst stain. Cells were visualized (20X magnification) (A) and % of γ-H2AX-positive cells was calculated (B) using Operetta high-content imaging and analysis system. Scale bars: 100 µm. (C and D) ARPE-19 cells were transfected with ATM, ATR or DNA-PK siRNAs or NT siRNA as a control. After 24 h, cells were infected with TB40/E-GFP-epi at 1 TCID50 per cell (A) or 100 μg/mL of zeocin (B). RNA samples were collected 24 h later. RT-qPCR was performed using specific primers to HSATII, ATM, ATR or DNA-PK transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (E) A-T(+) and A-T(-) cells were infected with TB40/E-GFP-epi at 1 TCID50 per cell, 200 μg/mL of zeocin or 200 μM of etoposide. RNA samples were collected 24 h later. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (F) ARPE-19 cells were treated with DMSO, 20 μM of either Ku-55933 or AZ31 for 2 h before zeocin was added at the concentration of 100 μg/mL. RNA samples were coll ected 24 hpt. RT-qPCR was performed using specific primers to HSATII, CDKN1A or DINO transcripts. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n =3. (G) ARPE-19 cells were treated with various concentrations of Ku-55933 or AZ31. At 24 hpt, cell viability was assessed. Data is presented as % viable cells, were averaged from at least three independent experiments and are presented as mean (SD). * P

    Article Snippet: DNA was RNase A treated (2 mg/ml; Qiagen) and viral DNA was quantified by qPCR using primers specific for the viral genomic UL44 (Forward: 5’-GTGCGCGCCCGATTTCAATATG-3’, Reverse: 5’-GCTTTCGCGCACAATGTCTTGG-3’) or cellular genomic GAPDH (Forward: 5’-CCCCACACACATGCACTTACC-3’, Reverse: 5’-CCTAGTCCCAGGGCTTTGATT-3’).

    Techniques: Expressing, Infection, Staining, Imaging, Transfection, Quantitative RT-PCR, Concentration Assay

    HSATII KD has strong effect on HCMV infection in epithelial cells. (A) RNA samples were collected at 24 hpi from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID50/cell. RT-qPCR was performed using HSATII-specific primers. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n=3. (B) Protein samples were collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. Protein levels were analyzed by the western blot technique using antibodies specific to IE1, IE2, pUL26, pUL44, pUL69, pp71 and pp28. Actin was used as a loading control. (C) Total DNA was collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID50/cell. vDNA and cellular DNA copy numbers were determined. Data are presented as a fold change mean ± SD of the relative vDNA to cellular DNA ratio. n=3 (D) Media samples were collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. TCID50 ml -1 values were determined . (E) Intracellular and extracellular virions were collected at indicated times from ARPE-19 cells transfected with either NT-LNA, HSATII-LNA#1, HSATII-LNA#2 or both HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. TCID50 ml -1 values were determined. Data are presented as a mean ± SD. n =3. *** P

    Journal: bioRxiv

    Article Title: HSATII RNA is induced via E2F3a and a non-canonical ATM-regulated DNA damage response pathway

    doi: 10.1101/2020.05.25.115238

    Figure Lengend Snippet: HSATII KD has strong effect on HCMV infection in epithelial cells. (A) RNA samples were collected at 24 hpi from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID50/cell. RT-qPCR was performed using HSATII-specific primers. GAPDH was used as an internal control. Data are presented as a fold change mean ± SD. n=3. (B) Protein samples were collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. Protein levels were analyzed by the western blot technique using antibodies specific to IE1, IE2, pUL26, pUL44, pUL69, pp71 and pp28. Actin was used as a loading control. (C) Total DNA was collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID50/cell. vDNA and cellular DNA copy numbers were determined. Data are presented as a fold change mean ± SD of the relative vDNA to cellular DNA ratio. n=3 (D) Media samples were collected at indicated times from ARPE-19 cells transfected with NT-LNA or HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. TCID50 ml -1 values were determined . (E) Intracellular and extracellular virions were collected at indicated times from ARPE-19 cells transfected with either NT-LNA, HSATII-LNA#1, HSATII-LNA#2 or both HSATII-LNAs 24 h before HCMV (TB40/E-GFP-epi) infection at 1 TCID 50 /cell. TCID50 ml -1 values were determined. Data are presented as a mean ± SD. n =3. *** P

    Article Snippet: DNA was RNase A treated (2 mg/ml; Qiagen) and viral DNA was quantified by qPCR using primers specific for the viral genomic UL44 (Forward: 5’-GTGCGCGCCCGATTTCAATATG-3’, Reverse: 5’-GCTTTCGCGCACAATGTCTTGG-3’) or cellular genomic GAPDH (Forward: 5’-CCCCACACACATGCACTTACC-3’, Reverse: 5’-CCTAGTCCCAGGGCTTTGATT-3’).

    Techniques: Infection, Transfection, Quantitative RT-PCR, Western Blot