polymerase chain reaction pcr system  (Thermo Fisher)


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    Structured Review

    Thermo Fisher polymerase chain reaction pcr system
    Polymerase Chain Reaction Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction pcr system/product/Thermo Fisher
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction pcr system - by Bioz Stars, 2020-04
    94/100 stars

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    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: The sample proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and were subsequently transferred from the gel onto a polyvinylidene difluoride (PVDF) membrane using an electric current. .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA).

    Amplification:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: .. The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems). ..

    Article Title: Association of Genetic Polymorphisms on VEGFA and VEGFR2 With Risk of Coronary Heart Disease
    Article Snippet: .. Tag SNPs were amplified with 9700 polymerase chain reaction (PCR) System (Applied Biosystems) with the primers designed by Primer 3 software (Table ). .. The PCR reaction mixture (10 μL) included 1 μL genomic DNA, 0.6 μL 5 μM designed primers, 0.4 μL 10 mM dNTP mix, 0.1 μL Taq DNA polymerase, 1 μL Taq DNA polymerase buffer, and 6.3 μL ultrapure water.

    Synthesized:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: Specific primers for CTGF/CCN2 were synthesized by Sigma Genosys (St. Louis, MO) (Forw. .. The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems).

    SYBR Green Assay:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: .. The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems). ..

    Cell Culture:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: A sample from a mouse ASC cultured on the plasma-treated and untreated PHB and PHBV films was lysed with RIPA lysis buffer. .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: .. The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems). ..

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA). .. Statistical Analysis The statistical analysis was performed using SPSS statistical software (SPSS version 23.0 for window, IBM Corporation, Selangor, Malaysia).

    Polymerase Chain Reaction:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: .. The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems). ..

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA). .. Statistical Analysis The statistical analysis was performed using SPSS statistical software (SPSS version 23.0 for window, IBM Corporation, Selangor, Malaysia).

    Article Title: Association of Genetic Polymorphisms on VEGFA and VEGFR2 With Risk of Coronary Heart Disease
    Article Snippet: .. Tag SNPs were amplified with 9700 polymerase chain reaction (PCR) System (Applied Biosystems) with the primers designed by Primer 3 software (Table ). .. The PCR reaction mixture (10 μL) included 1 μL genomic DNA, 0.6 μL 5 μM designed primers, 0.4 μL 10 mM dNTP mix, 0.1 μL Taq DNA polymerase, 1 μL Taq DNA polymerase buffer, and 6.3 μL ultrapure water.

    Incubation:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA). .. Statistical Analysis The statistical analysis was performed using SPSS statistical software (SPSS version 23.0 for window, IBM Corporation, Selangor, Malaysia).

    Western Blot:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: Paragraph title: 2.4.8. Western Blot Analysis ... The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Assembly of the Prothrombinase Complex on the Surface of Human Foreskin Fibroblasts: Implications for Connective Tissue Growth Factor
    Article Snippet: Paragraph title: RT-PCR analysis ... The cDNA was amplified, using Sybr green PCR master mix, by the quantitated Polymerase Chain Reaction (PCR) system (7500 real-time PCR system from Applied Biosystems).

    Lysis:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: A sample from a mouse ASC cultured on the plasma-treated and untreated PHB and PHBV films was lysed with RIPA lysis buffer. .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA).

    SDS Page:

    Article Title: Investigation and Characterization of Plasma-Treated Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymers for an In Vitro Cellular Study of Mouse Adipose-Derived Stem Cells
    Article Snippet: The sample proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and were subsequently transferred from the gel onto a polyvinylidene difluoride (PVDF) membrane using an electric current. .. The membrane was incubated with respective primary antibodies followed by secondary horseradish peroxidase-conjugated antibodies for 1 h. The protein signals were visualized using a polymerase chain reaction (PCR) system (Applied Biosystems StepOne PlusTM Real-Time PCR System Thermal; Foster City, CA, USA).

    Software:

    Article Title: Association of Genetic Polymorphisms on VEGFA and VEGFR2 With Risk of Coronary Heart Disease
    Article Snippet: .. Tag SNPs were amplified with 9700 polymerase chain reaction (PCR) System (Applied Biosystems) with the primers designed by Primer 3 software (Table ). .. The PCR reaction mixture (10 μL) included 1 μL genomic DNA, 0.6 μL 5 μM designed primers, 0.4 μL 10 mM dNTP mix, 0.1 μL Taq DNA polymerase, 1 μL Taq DNA polymerase buffer, and 6.3 μL ultrapure water.

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  • 95
    Thermo Fisher quan titative reverse transcriptase polymerase chain reaction qrt pcr
    miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by <t>qRT-PCR.</t> Values were normalized to GAPDH . (n=3; *P
    Quan Titative Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher sirna
    Knockdown of CacyBP facilitated viral replication and arrested apoptosis in DF-1 cells. DF-1 cells were transfected with mock and/or NC and si-CacyBP/SIP and incubated for 36 h. Mock, treated with <t>transfection</t> regent; NC, transfected with si-NC; si183/326/644, separately transfected with those siRNAs targets CacyBP/SIP. (A) Endogenous CacyBP/SIP was detected by western blotting. (B) DF-1 cells were co-transfected with si-CacyBP (326) and pCMV-HA-CacyBP. Thirty-six hours after transfection, protein expression of HA-CacyBP was detected by anti-HA antibody through immunofluorescence in DF-1 cells. (C) Replication kinetics of NDV RNAs from the mock, NC, and <t>siRNA</t> (326) groups of DF-1 cells; 1 MOI of F48E9 NDV were inoculated into the three groups of cells 24 h after transfection. Q-PCR was used to measure viral RNA replication 24 hpi. (D) Viral plaque formation tests were further used to measure the number of viruses in the supernatants. (E) Three cell cultures were prepared and transfected with mock, NC and siRNA (326), and then, the cells were washed and harvested, an annexin V assay was followed by flow cytometry to monitor for percentage of cells undergoing early apoptosis (bottom right quadrant) and late apoptosis(upper right quadrant). Y-axis is PI signal; X-asis is annexin V-FITC signal. Right graph data were the percentage of total apoptosis (bottom and upper right quadrant)and data from three independent experiments. (F–H) Q-PCR was used to analyze the expression of apoptosis-related markers and immune-associated markers 24 h after transfection with NC and si-CacyBP/SIP. (G) The transfected cell lysate was analyzed by western blotting with the indicated apoptosis-related antibody. Data shown in (C,D) are mean ± SD of four independent experiments, in (E,G,H) are mean ± SD of three independent expriments. * P
    Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt pcr rna
    MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by <t>RNA-immunoprecipitation</t> followed by <t>qRT-PCR.</t> Mean ± SD ( n = 3). P -value
    Qrt Pcr Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 285 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher human lung cancer a549
    Tetracenomycin X decreases the expression of cyclin D1 by the activation of p38 and c-JUN. ( A ) The <t>A549</t> and H460 cells were treated with tetracenomycin X for the indicated times or treated with various concentrations of tetracenomycin X for 8 h. The expression levels of the proteins were determined using western blotting. ( B ) The viability of the A5459 and H460 cells was determined after a 4-h pre-treatment with SP600125 or SB203580 and an 8-h treatment with tetracenomycin X. The expression levels of the proteins were determined using western blotting.
    Human Lung Cancer A549, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by qRT-PCR. Values were normalized to GAPDH . (n=3; *P

    Journal: International Journal of Molecular and Cellular Medicine

    Article Title: Overexpression of MiR-138 Inhibits Cell Growth and Induces Caspase-mediated Apoptosis in Acute Promyelocytic Leukemia Cell Line

    doi: 10.22088/IJMCM.BUMS.7.1.24

    Figure Lengend Snippet: miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by qRT-PCR. Values were normalized to GAPDH . (n=3; *P

    Article Snippet: The prepared cDNA was subjected to quan-titative reverse-transcriptase polymerase chain reaction (qRT-PCR), using Maxima SYBR Green Master mix (Thermo Scientific, Waltham, Massa-chusetts, USA) in the Rotor Gene 6000 Real Time PCR inst rument (Corbett Research, Hilden, Germany).

    Techniques: Transduction, Expressing, Quantitative RT-PCR

    Knockdown of CacyBP facilitated viral replication and arrested apoptosis in DF-1 cells. DF-1 cells were transfected with mock and/or NC and si-CacyBP/SIP and incubated for 36 h. Mock, treated with transfection regent; NC, transfected with si-NC; si183/326/644, separately transfected with those siRNAs targets CacyBP/SIP. (A) Endogenous CacyBP/SIP was detected by western blotting. (B) DF-1 cells were co-transfected with si-CacyBP (326) and pCMV-HA-CacyBP. Thirty-six hours after transfection, protein expression of HA-CacyBP was detected by anti-HA antibody through immunofluorescence in DF-1 cells. (C) Replication kinetics of NDV RNAs from the mock, NC, and siRNA (326) groups of DF-1 cells; 1 MOI of F48E9 NDV were inoculated into the three groups of cells 24 h after transfection. Q-PCR was used to measure viral RNA replication 24 hpi. (D) Viral plaque formation tests were further used to measure the number of viruses in the supernatants. (E) Three cell cultures were prepared and transfected with mock, NC and siRNA (326), and then, the cells were washed and harvested, an annexin V assay was followed by flow cytometry to monitor for percentage of cells undergoing early apoptosis (bottom right quadrant) and late apoptosis(upper right quadrant). Y-axis is PI signal; X-asis is annexin V-FITC signal. Right graph data were the percentage of total apoptosis (bottom and upper right quadrant)and data from three independent experiments. (F–H) Q-PCR was used to analyze the expression of apoptosis-related markers and immune-associated markers 24 h after transfection with NC and si-CacyBP/SIP. (G) The transfected cell lysate was analyzed by western blotting with the indicated apoptosis-related antibody. Data shown in (C,D) are mean ± SD of four independent experiments, in (E,G,H) are mean ± SD of three independent expriments. * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Newcastle Disease Virus V Protein Inhibits Cell Apoptosis and Promotes Viral Replication by Targeting CacyBP/SIP

    doi: 10.3389/fcimb.2018.00304

    Figure Lengend Snippet: Knockdown of CacyBP facilitated viral replication and arrested apoptosis in DF-1 cells. DF-1 cells were transfected with mock and/or NC and si-CacyBP/SIP and incubated for 36 h. Mock, treated with transfection regent; NC, transfected with si-NC; si183/326/644, separately transfected with those siRNAs targets CacyBP/SIP. (A) Endogenous CacyBP/SIP was detected by western blotting. (B) DF-1 cells were co-transfected with si-CacyBP (326) and pCMV-HA-CacyBP. Thirty-six hours after transfection, protein expression of HA-CacyBP was detected by anti-HA antibody through immunofluorescence in DF-1 cells. (C) Replication kinetics of NDV RNAs from the mock, NC, and siRNA (326) groups of DF-1 cells; 1 MOI of F48E9 NDV were inoculated into the three groups of cells 24 h after transfection. Q-PCR was used to measure viral RNA replication 24 hpi. (D) Viral plaque formation tests were further used to measure the number of viruses in the supernatants. (E) Three cell cultures were prepared and transfected with mock, NC and siRNA (326), and then, the cells were washed and harvested, an annexin V assay was followed by flow cytometry to monitor for percentage of cells undergoing early apoptosis (bottom right quadrant) and late apoptosis(upper right quadrant). Y-axis is PI signal; X-asis is annexin V-FITC signal. Right graph data were the percentage of total apoptosis (bottom and upper right quadrant)and data from three independent experiments. (F–H) Q-PCR was used to analyze the expression of apoptosis-related markers and immune-associated markers 24 h after transfection with NC and si-CacyBP/SIP. (G) The transfected cell lysate was analyzed by western blotting with the indicated apoptosis-related antibody. Data shown in (C,D) are mean ± SD of four independent experiments, in (E,G,H) are mean ± SD of three independent expriments. * P

    Article Snippet: Transfection and viral infection Plasmid or siRNA was transfected to DF-1 and/or Vero cells using Lipofectamine 2000 (Thermo Scientific, NH, USA) according to the manufacturer's instruction.

    Techniques: Transfection, Incubation, Western Blot, Expressing, Immunofluorescence, Polymerase Chain Reaction, Annexin V Assay, Flow Cytometry, Cytometry

    MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by RNA-immunoprecipitation followed by qRT-PCR. Mean ± SD ( n = 3). P -value

    Journal: Nature Communications

    Article Title: LOTUS domain protein MARF1 binds CCR4-NOT deadenylase complex to post-transcriptionally regulate gene expression in oocytes

    doi: 10.1038/s41467-018-06404-w

    Figure Lengend Snippet: MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by RNA-immunoprecipitation followed by qRT-PCR. Mean ± SD ( n = 3). P -value

    Article Snippet: qRT-PCR RNA from oocytes was prepared using miRVana (Thermo Fisher Scientific).

    Techniques: Immunoprecipitation, Western Blot, Expressing, Negative Control, Quantitative RT-PCR

    Tethered MARF1 shortens reporter mRNA poly-A tail and reduces reporter protein level. a GFP-5x BoxB reporter structure, harboring a ubiquitous tubulin promoter, GFP-coding sequence, and a 3′ UTR containing five BoxB hairpins. LambdaN-HA-fused control peptide, MARF1, GW182, and Piwi under a UASP promoter were expressed in germline cells using maternal-alpha-tubulin-Gal4 driver. b Confocal images of GFP signal in stage 14 oocytes. Scale bar is 100 μm. c Western blots using stage 14 oocyte lysates. Black triangles indicate the transgenic lambda-HA-fused proteins. d Quantification of band intensities in c . e ePAT assay measuring GFP reporter mRNA poly-A tail length in stage 14 oocytes. The amplified DNA sizes were analyzed on an agarose gel. f ePAT assay measuring poly-A tail lengths of GFP reporter mRNA and a negative control cdk1 mRNA in stage 14 oocytes. Amplified DNA sizes were analyzed by capillary fragment analysis. g Relative abundance of GFP-5xBoxB mRNA normalized by gapdh mRNA determined by qRT-PCR. Mean ± SD ( n = 6 for the control and n = 4 for all the others)

    Journal: Nature Communications

    Article Title: LOTUS domain protein MARF1 binds CCR4-NOT deadenylase complex to post-transcriptionally regulate gene expression in oocytes

    doi: 10.1038/s41467-018-06404-w

    Figure Lengend Snippet: Tethered MARF1 shortens reporter mRNA poly-A tail and reduces reporter protein level. a GFP-5x BoxB reporter structure, harboring a ubiquitous tubulin promoter, GFP-coding sequence, and a 3′ UTR containing five BoxB hairpins. LambdaN-HA-fused control peptide, MARF1, GW182, and Piwi under a UASP promoter were expressed in germline cells using maternal-alpha-tubulin-Gal4 driver. b Confocal images of GFP signal in stage 14 oocytes. Scale bar is 100 μm. c Western blots using stage 14 oocyte lysates. Black triangles indicate the transgenic lambda-HA-fused proteins. d Quantification of band intensities in c . e ePAT assay measuring GFP reporter mRNA poly-A tail length in stage 14 oocytes. The amplified DNA sizes were analyzed on an agarose gel. f ePAT assay measuring poly-A tail lengths of GFP reporter mRNA and a negative control cdk1 mRNA in stage 14 oocytes. Amplified DNA sizes were analyzed by capillary fragment analysis. g Relative abundance of GFP-5xBoxB mRNA normalized by gapdh mRNA determined by qRT-PCR. Mean ± SD ( n = 6 for the control and n = 4 for all the others)

    Article Snippet: qRT-PCR RNA from oocytes was prepared using miRVana (Thermo Fisher Scientific).

    Techniques: Sequencing, Western Blot, Transgenic Assay, Amplification, Agarose Gel Electrophoresis, Negative Control, Quantitative RT-PCR

    Tetracenomycin X decreases the expression of cyclin D1 by the activation of p38 and c-JUN. ( A ) The A549 and H460 cells were treated with tetracenomycin X for the indicated times or treated with various concentrations of tetracenomycin X for 8 h. The expression levels of the proteins were determined using western blotting. ( B ) The viability of the A5459 and H460 cells was determined after a 4-h pre-treatment with SP600125 or SB203580 and an 8-h treatment with tetracenomycin X. The expression levels of the proteins were determined using western blotting.

    Journal: Marine Drugs

    Article Title: Tetracenomycin X Exerts Antitumour Activity in Lung Cancer Cells through the Downregulation of Cyclin D1

    doi: 10.3390/md17010063

    Figure Lengend Snippet: Tetracenomycin X decreases the expression of cyclin D1 by the activation of p38 and c-JUN. ( A ) The A549 and H460 cells were treated with tetracenomycin X for the indicated times or treated with various concentrations of tetracenomycin X for 8 h. The expression levels of the proteins were determined using western blotting. ( B ) The viability of the A5459 and H460 cells was determined after a 4-h pre-treatment with SP600125 or SB203580 and an 8-h treatment with tetracenomycin X. The expression levels of the proteins were determined using western blotting.

    Article Snippet: Cell Culture The human lung cancer A549, H460, H157, HCC827 and H1975 cell lines were kept in our laboratory and maintained in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA) and penicillin (100 U/mL)/streptomycin (100 µg/mL) (National China Pharmaceutical Inc., Beijing, China) at 37 °C in a 5% CO2 incubator.

    Techniques: Expressing, Activation Assay, Western Blot

    The antitumour activity of tetracenomycin X is independent of apoptosis and autophagy. ( A , C ) The A549 and H460 cells were treated with tetracenomycin X (2.5 and 5 µmol/L) for 8 h. The expression levels of the proteins were determined using western blotting. ( B ) The A549 and H460 cells were treated with tetracenomycin X (10 µmol/L) for 24 h. Apoptosis was detected by annexin V-FITC/PI staining and flow cytometry analysis.

    Journal: Marine Drugs

    Article Title: Tetracenomycin X Exerts Antitumour Activity in Lung Cancer Cells through the Downregulation of Cyclin D1

    doi: 10.3390/md17010063

    Figure Lengend Snippet: The antitumour activity of tetracenomycin X is independent of apoptosis and autophagy. ( A , C ) The A549 and H460 cells were treated with tetracenomycin X (2.5 and 5 µmol/L) for 8 h. The expression levels of the proteins were determined using western blotting. ( B ) The A549 and H460 cells were treated with tetracenomycin X (10 µmol/L) for 24 h. Apoptosis was detected by annexin V-FITC/PI staining and flow cytometry analysis.

    Article Snippet: Cell Culture The human lung cancer A549, H460, H157, HCC827 and H1975 cell lines were kept in our laboratory and maintained in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA) and penicillin (100 U/mL)/streptomycin (100 µg/mL) (National China Pharmaceutical Inc., Beijing, China) at 37 °C in a 5% CO2 incubator.

    Techniques: Activity Assay, Expressing, Western Blot, Staining, Flow Cytometry, Cytometry

    Tetracenomycin X induces the proteasomal degradation of cyclin D1. ( A ) RT-PCR analysis of the gene expression of cyclin D1 in the A549 cells and H460 cells. ( B ) The A549 cells and H460 cells were pre-treated with MG132 at the indicated concentration for 2 h and then co-treated with tetracenomycin X (5 µmol/L) for 12 h. The A549 cells and H460 cells were pre-treated with DMSO or tetracenomycin X (5 µmol/L) and then co-treated with 10 μg/mL of cycloheximide (CHX) for the indicated times.

    Journal: Marine Drugs

    Article Title: Tetracenomycin X Exerts Antitumour Activity in Lung Cancer Cells through the Downregulation of Cyclin D1

    doi: 10.3390/md17010063

    Figure Lengend Snippet: Tetracenomycin X induces the proteasomal degradation of cyclin D1. ( A ) RT-PCR analysis of the gene expression of cyclin D1 in the A549 cells and H460 cells. ( B ) The A549 cells and H460 cells were pre-treated with MG132 at the indicated concentration for 2 h and then co-treated with tetracenomycin X (5 µmol/L) for 12 h. The A549 cells and H460 cells were pre-treated with DMSO or tetracenomycin X (5 µmol/L) and then co-treated with 10 μg/mL of cycloheximide (CHX) for the indicated times.

    Article Snippet: Cell Culture The human lung cancer A549, H460, H157, HCC827 and H1975 cell lines were kept in our laboratory and maintained in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA) and penicillin (100 U/mL)/streptomycin (100 µg/mL) (National China Pharmaceutical Inc., Beijing, China) at 37 °C in a 5% CO2 incubator.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Concentration Assay

    Tetracenomycin X-induced cell cycle arrest in the G0/G1 phase and decreased the expression levels of cell cycle-related proteins in the lung cancer cells. ( A ) the A549 and H460 cells were treated with tetracenomycin X (5 or 10 µmol/L) for 16 h. The cell cycle was detected by flow cytometry analysis. ( B ) The A549 and H460 cells were treated with tetracenomycin X for the indicated times or treated with various concentrations of tetracenomycin X for 8 h. The expression levels of proteins were determined using western blotting.

    Journal: Marine Drugs

    Article Title: Tetracenomycin X Exerts Antitumour Activity in Lung Cancer Cells through the Downregulation of Cyclin D1

    doi: 10.3390/md17010063

    Figure Lengend Snippet: Tetracenomycin X-induced cell cycle arrest in the G0/G1 phase and decreased the expression levels of cell cycle-related proteins in the lung cancer cells. ( A ) the A549 and H460 cells were treated with tetracenomycin X (5 or 10 µmol/L) for 16 h. The cell cycle was detected by flow cytometry analysis. ( B ) The A549 and H460 cells were treated with tetracenomycin X for the indicated times or treated with various concentrations of tetracenomycin X for 8 h. The expression levels of proteins were determined using western blotting.

    Article Snippet: Cell Culture The human lung cancer A549, H460, H157, HCC827 and H1975 cell lines were kept in our laboratory and maintained in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA) and penicillin (100 U/mL)/streptomycin (100 µg/mL) (National China Pharmaceutical Inc., Beijing, China) at 37 °C in a 5% CO2 incubator.

    Techniques: Expressing, Flow Cytometry, Cytometry, Western Blot

    Tetracenomycin X selectively inhibits the cell proliferation of lung cancer cells. ( A ) The structure of tetracenomycin X. ( B ) The proliferative activity of the five lung cancer cells and lung fibroblasts after being treated with tetracenomycin X and Adriamycin (0.1563, 0.3125, 0.625, 1.25, 2.5, 5, 10 and 15 µmol/L) for 24 h was assessed by sulforhodamine B (SRB) assay. The survival rates were calculated as a ratio compared with the control group (untreated cells). The values represent the mean ± SD of three independent samples. Each experiment was repeated three times under each condition. ( C ) The cell morphology of the A549 and H460 cells under a 4 *0.1 microscope after treatment with tetracenomycin X for 24 h.

    Journal: Marine Drugs

    Article Title: Tetracenomycin X Exerts Antitumour Activity in Lung Cancer Cells through the Downregulation of Cyclin D1

    doi: 10.3390/md17010063

    Figure Lengend Snippet: Tetracenomycin X selectively inhibits the cell proliferation of lung cancer cells. ( A ) The structure of tetracenomycin X. ( B ) The proliferative activity of the five lung cancer cells and lung fibroblasts after being treated with tetracenomycin X and Adriamycin (0.1563, 0.3125, 0.625, 1.25, 2.5, 5, 10 and 15 µmol/L) for 24 h was assessed by sulforhodamine B (SRB) assay. The survival rates were calculated as a ratio compared with the control group (untreated cells). The values represent the mean ± SD of three independent samples. Each experiment was repeated three times under each condition. ( C ) The cell morphology of the A549 and H460 cells under a 4 *0.1 microscope after treatment with tetracenomycin X for 24 h.

    Article Snippet: Cell Culture The human lung cancer A549, H460, H157, HCC827 and H1975 cell lines were kept in our laboratory and maintained in RPMI-1640 medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA) and penicillin (100 U/mL)/streptomycin (100 µg/mL) (National China Pharmaceutical Inc., Beijing, China) at 37 °C in a 5% CO2 incubator.

    Techniques: Activity Assay, Sulforhodamine B Assay, Microscopy