real-time quantitative pcr Search Results


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    LabCoat Live Introduction to qPCR and Absolute Quantification 1 week Learn the fundamentals of how to perform qPCR and the absolute quantification AQ method of analysis This course provides professional
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    97
    TaKaRa real time quantitative polymerase chain reaction pcr
    Confirmation of <t>RNA</t> and protein expression levels by real-time quantitative <t>PCR</t> and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p
    Real Time Quantitative Polymerase Chain Reaction Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 97/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative polymerase chain reaction pcr
    il25 −/− mice are highly susceptible to EAE. (A) Taqman <t>PCR</t> analysis of IL-25 <t>mRNA</t> levels in spinal cords from C57BL/6 WT normal and diseased versus il25 -LacZ-KI normal and diseased ( n = 6 mice per group). Data shown are representative of at least two experiments. ND, not detectable. (B) IL-25 mRNA expression in CNS-resident microglia. Brain leukocytes were isolated from bone marrow chimeras expressing the CD45.2 allotype on radiation-resistant host cells and the CD45.1 allotype on donor leukocytes. CD11b + CD45.2 lo CD45.1 neg resident microglia were purified from > 100 irradiation bone marrow chimeric naive or diseased (day 14) mice. Data from quantitative real-time PCR analysis for IL-25 expression were normalized to ubiquitin. (C) Clinical score (top) and percentage of incidence of disease (bottom) of C57BL/6 (WT) and il25 −/− mice immunized with MOG/CFA s.c. on day 0 and PTX i.v. on days 0 and 2. The dagger indicates that by day 17, all il25 −/− mice had died from EAE-associated complications. Data shown are 7–9 mice per group and are representative of at least three experiments.
    Real Time Quantitative Polymerase Chain Reaction Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 914 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PCR Biosystems Ltd real time quantitative polymerase chain reaction pcr
    il25 −/− mice are highly susceptible to EAE. (A) Taqman <t>PCR</t> analysis of IL-25 <t>mRNA</t> levels in spinal cords from C57BL/6 WT normal and diseased versus il25 -LacZ-KI normal and diseased ( n = 6 mice per group). Data shown are representative of at least two experiments. ND, not detectable. (B) IL-25 mRNA expression in CNS-resident microglia. Brain leukocytes were isolated from bone marrow chimeras expressing the CD45.2 allotype on radiation-resistant host cells and the CD45.1 allotype on donor leukocytes. CD11b + CD45.2 lo CD45.1 neg resident microglia were purified from > 100 irradiation bone marrow chimeric naive or diseased (day 14) mice. Data from quantitative real-time PCR analysis for IL-25 expression were normalized to ubiquitin. (C) Clinical score (top) and percentage of incidence of disease (bottom) of C57BL/6 (WT) and il25 −/− mice immunized with MOG/CFA s.c. on day 0 and PTX i.v. on days 0 and 2. The dagger indicates that by day 17, all il25 −/− mice had died from EAE-associated complications. Data shown are 7–9 mice per group and are representative of at least three experiments.
    Real Time Quantitative Polymerase Chain Reaction Pcr, supplied by PCR Biosystems Ltd, 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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    Roche real time quantitative polymerase chain reaction real time quantitative pcr qpcr
    HT extracts inhibit gene expression. (a–c) Cells were treated with HT extracts at the indicated concentrations and quantitative <t>PCR</t> was performed ( n = 6/group). Data are expressed as means ± SD. ∗ P
    Real Time Quantitative Polymerase Chain Reaction Real Time Quantitative Pcr Qpcr, supplied by Roche, used in various techniques. Bioz Stars score: 77/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative polymerase chain reaction qrt pcr
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Real Time Quantitative Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 506 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time fluorescent polymerase chain reaction pcr
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Quantitative Real Time Fluorescent Polymerase Chain Reaction Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time fluorescence quantitative polymerase chain reaction pcr kit
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Real Time Fluorescence Quantitative Polymerase Chain Reaction Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche real time quantitative polymerase chain reaction rt pcr assay
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Real Time Quantitative Polymerase Chain Reaction Rt Pcr Assay, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene real time quantitative pcr qpcr
    p63 is significantly down-regulated in UM cell lines. ( A ) Representative western blot showing relative protein levels of endogenous p63 protein in UM cell lysates. Skin cell lysate served as a positive control for p63 antibody. ( B ) mRNA was extracted from non-transfected (NT) OCM-1 as well as from A431 cells and p63-tGFP-transfected OCM-1 cells used as positive controls for p63 expression. Quantitative <t>PCR</t> was performed to assess mRNA levels by analysing the amplification plots and dissociation curves for each of the three types of cells. NTC, non-template control.
    Real Time Quantitative Pcr Qpcr, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time quantitative polymerase chain reaction rt pcr kits
    p63 is significantly down-regulated in UM cell lines. ( A ) Representative western blot showing relative protein levels of endogenous p63 protein in UM cell lysates. Skin cell lysate served as a positive control for p63 antibody. ( B ) mRNA was extracted from non-transfected (NT) OCM-1 as well as from A431 cells and p63-tGFP-transfected OCM-1 cells used as positive controls for p63 expression. Quantitative <t>PCR</t> was performed to assess mRNA levels by analysing the amplification plots and dissociation curves for each of the three types of cells. NTC, non-template control.
    Real Time Quantitative Polymerase Chain Reaction Rt Pcr Kits, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative pcr qpcr
    Real-time quantitative <t>PCR</t> of PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) and anaerobic glycolysis genes during submergence. (a.) The expression of an unannotated PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) gene in submerged shoot tissue during submergence. (b.) PYRUVATE DECARBOXYLASE3 ( PDC3 ) expression in shoot tissues during submergence. (c.) ALCOHOL DEHYDROGENASE1 ( ADH1 ) transcript levels in submerged shoot tissue. All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p
    Real Time Quantitative Pcr Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2609 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies real time quantitative pcr qpcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Real Time Quantitative Pcr Qpcr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quanta Biosciences real time quantitative pcr qpcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Real Time Quantitative Pcr Qpcr, supplied by Quanta Biosciences, used in various techniques. Bioz Stars score: 90/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher stepone real time quantitative polymerase chain reaction qrt pcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Stepone Real Time Quantitative Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eppendorf AG real time quantitative pcr qpcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Real Time Quantitative Pcr Qpcr, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad quantitative real time polymerase chain reaction qpcr quantitative real time pcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Quantitative Real Time Polymerase Chain Reaction Qpcr Quantitative Real Time Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies real time quantitative polymerase chain reaction pcr assay real time quantitative pcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Real Time Quantitative Polymerase Chain Reaction Pcr Assay Real Time Quantitative Pcr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher thermo script real time quantitative polymerase chain reaction rt pcr
    <t>PCR</t> and <t>qPCR</t> validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.
    Thermo Script Real Time Quantitative Polymerase Chain Reaction Rt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time quantitative polymerase chain reaction qrt pcr real time quantitative pcr
    Validation of the accuracy of RNA-seq data. Note: The validation of the accuracy of RNA-seq data was carried out by comparing of RPKM and the relative expression of five randomly selected genes using <t>qRT-PCR</t> for each tissue and time point.
    Real Time Quantitative Polymerase Chain Reaction Qrt Pcr Real Time Quantitative Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche multiplex real time quantitative polymerase chain reaction pcr technology
    Validation of the accuracy of RNA-seq data. Note: The validation of the accuracy of RNA-seq data was carried out by comparing of RPKM and the relative expression of five randomly selected genes using <t>qRT-PCR</t> for each tissue and time point.
    Multiplex Real Time Quantitative Polymerase Chain Reaction Pcr Technology, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi 7900 real time quantitative polymerase chain reaction pcr instrument
    Validation of the accuracy of RNA-seq data. Note: The validation of the accuracy of RNA-seq data was carried out by comparing of RPKM and the relative expression of five randomly selected genes using <t>qRT-PCR</t> for each tissue and time point.
    Abi 7900 Real Time Quantitative Polymerase Chain Reaction Pcr Instrument, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega real time fluorescence quantitative polymerase chain reaction qrt pcr kit
    Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by <t>qRT-PCR,</t> which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P
    Real Time Fluorescence Quantitative Polymerase Chain Reaction Qrt Pcr Kit, supplied by Promega, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene real time quantitative pcr
    Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by <t>qRT-PCR,</t> which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P
    Real Time Quantitative Pcr, supplied by OriGene, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The inhibitors 17-AAG, EC154, and LBH589 inhibit HIF-dependent gene transcription in CCRCC cells . The CCRCC cell line UMRC2 was treated for 16 h with inhibitors as in Figure 1. Total mRNA was isolated and <t>HIF-α</t> regulated genes analyzed by <t>qRT-PCR.</t> Values were normalized to GAPDH and are presented relative to control cells, with standard deviation shown. All drug treatments significantly reduced all transcript levels (*) as determined by ANOVA and Student's t- test ( p
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    The inhibitors 17-AAG, EC154, and LBH589 inhibit HIF-dependent gene transcription in CCRCC cells . The CCRCC cell line UMRC2 was treated for 16 h with inhibitors as in Figure 1. Total mRNA was isolated and <t>HIF-α</t> regulated genes analyzed by <t>qRT-PCR.</t> Values were normalized to GAPDH and are presented relative to control cells, with standard deviation shown. All drug treatments significantly reduced all transcript levels (*) as determined by ANOVA and Student's t- test ( p
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    Effect of labeling on chondrogenic differentiation of ASC. (A) Unlabeled, (B) BNF starch- and (C, E, F) nanomag-D-spio-labeled cells were chondrogenically stimulated for 21 d. Pellets were analyzed using Heidenhain's AZAN trichrome staining showing a reduced collagen type II-positive extracellular matrix (light blue) due to nanoparticle labeling. BNF-labeled cells (25 and 50 µg Fe/ml) failed to generate compact pellets (not shown) (Axio Imager M2, Carl Zeiss Microscopy GmbH, Jena, Germany; scale bars = 100 µm). (D) Collagen type II was analyzed using Real-Time quantitative <t>PCR</t> revealing a diminished mRNA expression of collagen type II due to nanoparticle labeling (2∧ΔΔCT ± %CV; n = 2, normalized to β-actin).
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    Effect of labeling on chondrogenic differentiation of ASC. (A) Unlabeled, (B) BNF starch- and (C, E, F) nanomag-D-spio-labeled cells were chondrogenically stimulated for 21 d. Pellets were analyzed using Heidenhain's AZAN trichrome staining showing a reduced collagen type II-positive extracellular matrix (light blue) due to nanoparticle labeling. BNF-labeled cells (25 and 50 µg Fe/ml) failed to generate compact pellets (not shown) (Axio Imager M2, Carl Zeiss Microscopy GmbH, Jena, Germany; scale bars = 100 µm). (D) Collagen type II was analyzed using Real-Time quantitative <t>PCR</t> revealing a diminished mRNA expression of collagen type II due to nanoparticle labeling (2∧ΔΔCT ± %CV; n = 2, normalized to β-actin).
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    Magnitude of oval cell response quantified by real time <t>PCR</t> analysis of AFP <t>mRNA,</t> relative to normal liver tissue expression and normalized to beta actin (A), 9AP – animals fed normal rat chow and subjected to 2AAF/PH protocol; 9DAP – animals maintained on the L-cysteine diet associated to 2AAF/PH treatment. GAPDH was used as a loading control (B).
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    Magnitude of oval cell response quantified by real time <t>PCR</t> analysis of AFP <t>mRNA,</t> relative to normal liver tissue expression and normalized to beta actin (A), 9AP – animals fed normal rat chow and subjected to 2AAF/PH protocol; 9DAP – animals maintained on the L-cysteine diet associated to 2AAF/PH treatment. GAPDH was used as a loading control (B).
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    Magnitude of oval cell response quantified by real time <t>PCR</t> analysis of AFP <t>mRNA,</t> relative to normal liver tissue expression and normalized to beta actin (A), 9AP – animals fed normal rat chow and subjected to 2AAF/PH protocol; 9DAP – animals maintained on the L-cysteine diet associated to 2AAF/PH treatment. GAPDH was used as a loading control (B).
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    Shanghai Kehua real time quantitative pcr
    The GM-CSF/IFN-α/VACCINE promoted the clearance of HBcAg-positive hepatocytes ( A ) 14 days after the fourth immunization, liver sections were stained for HBcAg (brown staining) by IHC. Scale bar represents 50 μm. ( B ) 14 days after the fourth immunization, the <t>HBV</t> DNA levels in liver were analyzed by <t>q-PCR.</t> ( C – D ) 14 days after the fourth immunization, serum ALT was measured and the liver-infiltrating lymphocytes were stained by H E. Scale bar represents 50 μm. Symbols represent mean ± SEM. * P
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    Image Search Results


    Confirmation of RNA and protein expression levels by real-time quantitative PCR and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p

    Journal: Proteome Science

    Article Title: Proteomic analysis on effectors involved in BMP-2-induced osteogenic differentiation of beagle bone marrow mesenchymal stem cells

    doi: 10.1186/1477-5956-12-13

    Figure Lengend Snippet: Confirmation of RNA and protein expression levels by real-time quantitative PCR and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p

    Article Snippet: Real-time quantitative PCR The RNA extracted from BMSCs (1 μg) was first reverse-transcribed into cDNA (PrimeScript; Takara, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    il25 −/− mice are highly susceptible to EAE. (A) Taqman PCR analysis of IL-25 mRNA levels in spinal cords from C57BL/6 WT normal and diseased versus il25 -LacZ-KI normal and diseased ( n = 6 mice per group). Data shown are representative of at least two experiments. ND, not detectable. (B) IL-25 mRNA expression in CNS-resident microglia. Brain leukocytes were isolated from bone marrow chimeras expressing the CD45.2 allotype on radiation-resistant host cells and the CD45.1 allotype on donor leukocytes. CD11b + CD45.2 lo CD45.1 neg resident microglia were purified from > 100 irradiation bone marrow chimeric naive or diseased (day 14) mice. Data from quantitative real-time PCR analysis for IL-25 expression were normalized to ubiquitin. (C) Clinical score (top) and percentage of incidence of disease (bottom) of C57BL/6 (WT) and il25 −/− mice immunized with MOG/CFA s.c. on day 0 and PTX i.v. on days 0 and 2. The dagger indicates that by day 17, all il25 −/− mice had died from EAE-associated complications. Data shown are 7–9 mice per group and are representative of at least three experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: IL-25 regulates Th17 function in autoimmune inflammation

    doi: 10.1084/jem.20061738

    Figure Lengend Snippet: il25 −/− mice are highly susceptible to EAE. (A) Taqman PCR analysis of IL-25 mRNA levels in spinal cords from C57BL/6 WT normal and diseased versus il25 -LacZ-KI normal and diseased ( n = 6 mice per group). Data shown are representative of at least two experiments. ND, not detectable. (B) IL-25 mRNA expression in CNS-resident microglia. Brain leukocytes were isolated from bone marrow chimeras expressing the CD45.2 allotype on radiation-resistant host cells and the CD45.1 allotype on donor leukocytes. CD11b + CD45.2 lo CD45.1 neg resident microglia were purified from > 100 irradiation bone marrow chimeric naive or diseased (day 14) mice. Data from quantitative real-time PCR analysis for IL-25 expression were normalized to ubiquitin. (C) Clinical score (top) and percentage of incidence of disease (bottom) of C57BL/6 (WT) and il25 −/− mice immunized with MOG/CFA s.c. on day 0 and PTX i.v. on days 0 and 2. The dagger indicates that by day 17, all il25 −/− mice had died from EAE-associated complications. Data shown are 7–9 mice per group and are representative of at least three experiments.

    Article Snippet: Cytokine-specific mRNA was measured by real-time quantitative PCR (Applied Biosystems).

    Techniques: Mouse Assay, Polymerase Chain Reaction, Expressing, Isolation, Purification, Irradiation, Real-time Polymerase Chain Reaction

    IL-13 is required for Th17 suppression in IL-25–mediated protection from EAE. Clinical score of il4 −/− (A), il13 −/− (B), or il4ra −/− mice (C), with five mice per group injected daily with PBS or mIL-25 (s.c.) from days 1–12. Results are representative of two experiments. (D–F) Naive T cells and LPS-activated CD11c + DCs were obtained from DO11.10 × RAG −/− mice (pool of five mice) and incubated for 3 d with IL-23 in the presence of OVA peptide. IFNγ and IL-4 were neutralized in these cultures. (D) The effect of IL-25 addition on the number of IL-23–induced IL-17 producers was assessed by intracellular staining for IL-17. (E) IL-13 expression in these cultures was determined by LUMINEX. (F) The modulation of IL-17 protein production in IL-23–driven cultures by IL-25, IL-25 plus anti–IL-13, and IL-13 is shown in percentage of IL-23–induced IL-17 production without addition of exogenous factors (mean IL-17 production is 202.3 pg/ml, assessed by LUMINEX). Data shown are representative of at least two experiments (D and E) or pooled data from two similar experiments (F). (G) Purified MOG 33-55 TCR transgenic T cells and LPS-activated CD11c + DCs were incubated for 4 d with IL-23 in the presence of MOG peptide and neutralizing antibodies against IFNγ and IL-4. IL-17 production after incubation with IL-25, IL-25 plus anti–IL-13, and IL-13 was measured by LUMINEX. (H) DCs were purified from spleens and superficial lymph nodes and activated with LPS with or without addition of IL-13. After 24 h, mRNA expression relative to ubiquitin was measured for IL-23p19, IL-1β, and IL-6 by quantitative real-time PCR. Shown is an experiment using DCs from C57BL/6 mice representative of two C57BL/6 and two BALB/c experiments with a similar outcome ( n = 3–5 mice pooled per experiment).

    Journal: The Journal of Experimental Medicine

    Article Title: IL-25 regulates Th17 function in autoimmune inflammation

    doi: 10.1084/jem.20061738

    Figure Lengend Snippet: IL-13 is required for Th17 suppression in IL-25–mediated protection from EAE. Clinical score of il4 −/− (A), il13 −/− (B), or il4ra −/− mice (C), with five mice per group injected daily with PBS or mIL-25 (s.c.) from days 1–12. Results are representative of two experiments. (D–F) Naive T cells and LPS-activated CD11c + DCs were obtained from DO11.10 × RAG −/− mice (pool of five mice) and incubated for 3 d with IL-23 in the presence of OVA peptide. IFNγ and IL-4 were neutralized in these cultures. (D) The effect of IL-25 addition on the number of IL-23–induced IL-17 producers was assessed by intracellular staining for IL-17. (E) IL-13 expression in these cultures was determined by LUMINEX. (F) The modulation of IL-17 protein production in IL-23–driven cultures by IL-25, IL-25 plus anti–IL-13, and IL-13 is shown in percentage of IL-23–induced IL-17 production without addition of exogenous factors (mean IL-17 production is 202.3 pg/ml, assessed by LUMINEX). Data shown are representative of at least two experiments (D and E) or pooled data from two similar experiments (F). (G) Purified MOG 33-55 TCR transgenic T cells and LPS-activated CD11c + DCs were incubated for 4 d with IL-23 in the presence of MOG peptide and neutralizing antibodies against IFNγ and IL-4. IL-17 production after incubation with IL-25, IL-25 plus anti–IL-13, and IL-13 was measured by LUMINEX. (H) DCs were purified from spleens and superficial lymph nodes and activated with LPS with or without addition of IL-13. After 24 h, mRNA expression relative to ubiquitin was measured for IL-23p19, IL-1β, and IL-6 by quantitative real-time PCR. Shown is an experiment using DCs from C57BL/6 mice representative of two C57BL/6 and two BALB/c experiments with a similar outcome ( n = 3–5 mice pooled per experiment).

    Article Snippet: Cytokine-specific mRNA was measured by real-time quantitative PCR (Applied Biosystems).

    Techniques: Mouse Assay, Injection, Incubation, Staining, Expressing, Luminex, Purification, Transgenic Assay, Real-time Polymerase Chain Reaction

    HT extracts inhibit gene expression. (a–c) Cells were treated with HT extracts at the indicated concentrations and quantitative PCR was performed ( n = 6/group). Data are expressed as means ± SD. ∗ P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Helicteric Acid, Oleanic Acid, and Betulinic Acid, Three Triterpenes from Helicteres angustifolia L., Inhibit Proliferation and Induce Apoptosis in HT-29 Colorectal Cancer Cells via Suppressing NF-κB and STAT3 Signaling

    doi: 10.1155/2017/5180707

    Figure Lengend Snippet: HT extracts inhibit gene expression. (a–c) Cells were treated with HT extracts at the indicated concentrations and quantitative PCR was performed ( n = 6/group). Data are expressed as means ± SD. ∗ P

    Article Snippet: Real-time quantitative PCR (qPCR) was performed to quantify mRNA levels using the SYBR-Green PCR kit (Roche, Indianapolis, IN, USA) on the LightCycler 480 Real-Time PCR System (Roche).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Isolated around-the-clock α- and β-cells express in vivo circadian rhythm for selected core clock genes after fluorescence-activated cell sorting (FACS) separation . Triple transgenic mice (ProGcg-Venus/RIP-Cherry/Per2:Luc) were kept at standard 12 h/12 h light–dark cycles (ZT0 (Zeitgeber) corresponds to 07:00 a.m., and ZT12 corresponds to 07:00 p.m.), with constant access to the drinking water and night-restricted feeding regime for 2 weeks prior to islet collection. Pancreatic islets were isolated around-the-clock every 4 h and subjected to the FACS separation procedure. Temporal profiles of selected core clock transcripts ( Clock, Per1 , and Per2 ) were assessed in freshly extracted mRNA at each time point from α- and β-cell populations by quantitative RT-PCR analysis. Transcript levels were normalized to Hprt expression. Data expressed as mean ± SD ( N = 2 biological replicates of 6 mice at each time point).

    Journal: Frontiers in Endocrinology

    Article Title: High-Resolution Recording of the Circadian Oscillator in Primary Mouse α- and β-Cell Culture

    doi: 10.3389/fendo.2017.00068

    Figure Lengend Snippet: Isolated around-the-clock α- and β-cells express in vivo circadian rhythm for selected core clock genes after fluorescence-activated cell sorting (FACS) separation . Triple transgenic mice (ProGcg-Venus/RIP-Cherry/Per2:Luc) were kept at standard 12 h/12 h light–dark cycles (ZT0 (Zeitgeber) corresponds to 07:00 a.m., and ZT12 corresponds to 07:00 p.m.), with constant access to the drinking water and night-restricted feeding regime for 2 weeks prior to islet collection. Pancreatic islets were isolated around-the-clock every 4 h and subjected to the FACS separation procedure. Temporal profiles of selected core clock transcripts ( Clock, Per1 , and Per2 ) were assessed in freshly extracted mRNA at each time point from α- and β-cell populations by quantitative RT-PCR analysis. Transcript levels were normalized to Hprt expression. Data expressed as mean ± SD ( N = 2 biological replicates of 6 mice at each time point).

    Article Snippet: Specific target gene mRNA levels were analyzed by real-time quantitative PCR using the LightCycler technology (LC480; Roche Diagnostics).

    Techniques: Isolation, In Vivo, Fluorescence, FACS, Transgenic Assay, Mouse Assay, Quantitative RT-PCR, Expressing

    Effect of selective retinoic acid receptor (RAR) agonists on interleukin-1 (IL-1)-induced expression of IL-6 . (a) To control the ability of the selective agonists to trigger RAR-dependent responses, rat synovial fibroblasts were stimulated for 6 hours with 0.1 μM of RAR agonist (BMS-753 for RAR-α, BMS-453 for RAR-β, or BMS-961 for RAR-γ), and the mRNA level of the target gene RAR-β normalized to ribosomal protein S29 ( RP29 ) was studied by real-time polymerase chain reaction (PCR). (b) The suppressive effect of RAR-selective agonists on the IL-6 mRNA level normalized to RP29 was studied by real-time PCR in cells stimulated with 10 ng/mL of IL-1β in the presence or absence of 0.1 or 1 μM of RAR agonist. Data are expressed as mean ± standard deviation of values from at least three independent experiments. Statistically significant differences from the control are indicated as * P

    Journal: Arthritis Research & Therapy

    Article Title: All-trans retinoic acid suppresses interleukin-6 expression in interleukin-1-stimulated synovial fibroblasts by inhibition of ERK1/2 pathway independently of RAR activation

    doi: 10.1186/ar2569

    Figure Lengend Snippet: Effect of selective retinoic acid receptor (RAR) agonists on interleukin-1 (IL-1)-induced expression of IL-6 . (a) To control the ability of the selective agonists to trigger RAR-dependent responses, rat synovial fibroblasts were stimulated for 6 hours with 0.1 μM of RAR agonist (BMS-753 for RAR-α, BMS-453 for RAR-β, or BMS-961 for RAR-γ), and the mRNA level of the target gene RAR-β normalized to ribosomal protein S29 ( RP29 ) was studied by real-time polymerase chain reaction (PCR). (b) The suppressive effect of RAR-selective agonists on the IL-6 mRNA level normalized to RP29 was studied by real-time PCR in cells stimulated with 10 ng/mL of IL-1β in the presence or absence of 0.1 or 1 μM of RAR agonist. Data are expressed as mean ± standard deviation of values from at least three independent experiments. Statistically significant differences from the control are indicated as * P

    Article Snippet: The mRNA levels for RAR-α , -β , or -γ , RXR-α , -β , or -γ , IL-6 , and ribosomal protein S29 (RP29 ) were quantified by real-time quantitative polymerase chain reaction (PCR) in capillaries with the Lightcycler™ technology (Roche Molecular Biochemicals).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Standard Deviation

    QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Amplification

    QRT-PCR for 27-hydroxylase and ABCA1 message in indomethacin-treated THP-1 cells. (a) 27-Hydroxylase mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. At 50 mM indomethacin concentration, cell death was statistically significant (16.8% ± 1.0%). ABCA1, ATP-binding cassette transporter A1; COX, cyclooxygenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; QRT-PCR, quantitative real-time polymerase chain reaction. * p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in indomethacin-treated THP-1 cells. (a) 27-Hydroxylase mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. At 50 mM indomethacin concentration, cell death was statistically significant (16.8% ± 1.0%). ABCA1, ATP-binding cassette transporter A1; COX, cyclooxygenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; QRT-PCR, quantitative real-time polymerase chain reaction. * p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Isolation, Amplification, Concentration Assay, Binding Assay, Real-time Polymerase Chain Reaction

    QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 macrophages exposed to prostaglandins or TXA 2 . (a) 27-Hydroxylase message in THP-1 macrophages is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 in THP-1 macrophages and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. * p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 macrophages exposed to prostaglandins or TXA 2 . (a) 27-Hydroxylase message in THP-1 macrophages is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 in THP-1 macrophages and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. * p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Amplification

    p63 is significantly down-regulated in UM cell lines. ( A ) Representative western blot showing relative protein levels of endogenous p63 protein in UM cell lysates. Skin cell lysate served as a positive control for p63 antibody. ( B ) mRNA was extracted from non-transfected (NT) OCM-1 as well as from A431 cells and p63-tGFP-transfected OCM-1 cells used as positive controls for p63 expression. Quantitative PCR was performed to assess mRNA levels by analysing the amplification plots and dissociation curves for each of the three types of cells. NTC, non-template control.

    Journal: British Journal of Cancer

    Article Title: p63 is required beside p53 for PERP-mediated apoptosis in uveal melanoma

    doi: 10.1038/bjc.2016.269

    Figure Lengend Snippet: p63 is significantly down-regulated in UM cell lines. ( A ) Representative western blot showing relative protein levels of endogenous p63 protein in UM cell lysates. Skin cell lysate served as a positive control for p63 antibody. ( B ) mRNA was extracted from non-transfected (NT) OCM-1 as well as from A431 cells and p63-tGFP-transfected OCM-1 cells used as positive controls for p63 expression. Quantitative PCR was performed to assess mRNA levels by analysing the amplification plots and dissociation curves for each of the three types of cells. NTC, non-template control.

    Article Snippet: Real-time quantitative PCR (qPCR) was performed with gene-specific primer sets for PERP, p63 and GAPDH ( ) using a Stratagene MX3000P qPCR System (Stratagene, La Jolla, CA, USA) and MESA Blue qPCR Kit for SYBR Green assay (Eurogentec, Southampton, UK) according to the manufacturer's instructions.

    Techniques: Western Blot, Positive Control, Transfection, Expressing, Real-time Polymerase Chain Reaction, Amplification

    miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis. ( A ) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and ( B ) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 ( C ) and NCI-H1299 ( D ) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 ( E , G ) and NCI-H1299 ( F , H ) cells. * P

    Journal: Scientific Reports

    Article Title: MircoRNA-33a inhibits epithelial-to-mesenchymal transition and metastasis and could be a prognostic marker in non-small cell lung cancer

    doi: 10.1038/srep13677

    Figure Lengend Snippet: miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis. ( A ) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and ( B ) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 ( C ) and NCI-H1299 ( D ) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 ( E , G ) and NCI-H1299 ( F , H ) cells. * P

    Article Snippet: For the miR-33a quantification, real-time quantitative PCR of miR-33a was performed on a Mx3005P system (Stratagene, USA) using the Hairpin-itTM miR qPCR quantification kit (GenePharma, China) and the SYBR Premix ExTaq II reagent (Takara, Japan).

    Techniques: Over Expression, Transfection, Quantitative RT-PCR, Western Blot, Biomarker Assay, Migration

    Low-expression of miR-33a predicts poor prognosis in NSCLC patients. ( A ) Relative miR-33a expression levels were measured in 53 pairs of NSCLC tumor and non-tumor tissue samples by real-time quantitative RT-PCR. ( B ) The association of local invasion and overall survival in NSCLC patients. ( C ) The expression level of miR-33a in relation to overall survival in NSCLC patients. *** P

    Journal: Scientific Reports

    Article Title: MircoRNA-33a inhibits epithelial-to-mesenchymal transition and metastasis and could be a prognostic marker in non-small cell lung cancer

    doi: 10.1038/srep13677

    Figure Lengend Snippet: Low-expression of miR-33a predicts poor prognosis in NSCLC patients. ( A ) Relative miR-33a expression levels were measured in 53 pairs of NSCLC tumor and non-tumor tissue samples by real-time quantitative RT-PCR. ( B ) The association of local invasion and overall survival in NSCLC patients. ( C ) The expression level of miR-33a in relation to overall survival in NSCLC patients. *** P

    Article Snippet: For the miR-33a quantification, real-time quantitative PCR of miR-33a was performed on a Mx3005P system (Stratagene, USA) using the Hairpin-itTM miR qPCR quantification kit (GenePharma, China) and the SYBR Premix ExTaq II reagent (Takara, Japan).

    Techniques: Expressing, Quantitative RT-PCR

    Real-time quantitative PCR of PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) and anaerobic glycolysis genes during submergence. (a.) The expression of an unannotated PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) gene in submerged shoot tissue during submergence. (b.) PYRUVATE DECARBOXYLASE3 ( PDC3 ) expression in shoot tissues during submergence. (c.) ALCOHOL DEHYDROGENASE1 ( ADH1 ) transcript levels in submerged shoot tissue. All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Journal: PLoS ONE

    Article Title: Genetic and Molecular Characterization of Submergence Response Identifies Subtol6 as a Major Submergence Tolerance Locus in Maize

    doi: 10.1371/journal.pone.0120385

    Figure Lengend Snippet: Real-time quantitative PCR of PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) and anaerobic glycolysis genes during submergence. (a.) The expression of an unannotated PYROPHOSPHATE-DEPENDANT FRUCTOSE-6-PHOSPHATE 1-PHOSPHOTRANSERASE ( PFP ) gene in submerged shoot tissue during submergence. (b.) PYRUVATE DECARBOXYLASE3 ( PDC3 ) expression in shoot tissues during submergence. (c.) ALCOHOL DEHYDROGENASE1 ( ADH1 ) transcript levels in submerged shoot tissue. All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Article Snippet: cDNA synthesis and quantitative PCR First strand cDNA synthesis was performed using SuperScript VILO for real-time quantitative PCR (qPCR) (Invitrogen Corp., Carlsbad, CA, USA).

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Detection of ROS and expression of ROS marker genes in inbreds with contrasting levels of submergence tolerance. (a.) Detection of H 2 O 2 and superoxide in selected submergence tolerant and sensitive maize genotypes. A subset of four nested association mapping panel parents were selected based on visual leaf scoring for ROS staining assays after submergence. Hydrogen peroxide was detected using stain 3,3′-diaminobenzidine tetrahydrochloride (DAB), while superoxide was detected using nitroblue tetrazolium (NBT). Both assays were performed after 96 h of submergence. In B97, the third leaf was barely emerged and not used for the assay. (b-d) Real-time quantitative PCR of ROS genes. ROS genes were selected based on the study by Gadjev, Vanderauwera and Gechev [ 30 ] and showed a significant induction in response to at least two stresses that induce ROS formation in Arabidopsis. (b.) ALTERNATIVE OXIDASE 1a ( AOX1a ); (c.) WRKY6 (d.) CYTOCHROME P81 D8 ( CYP81D8 ). All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Journal: PLoS ONE

    Article Title: Genetic and Molecular Characterization of Submergence Response Identifies Subtol6 as a Major Submergence Tolerance Locus in Maize

    doi: 10.1371/journal.pone.0120385

    Figure Lengend Snippet: Detection of ROS and expression of ROS marker genes in inbreds with contrasting levels of submergence tolerance. (a.) Detection of H 2 O 2 and superoxide in selected submergence tolerant and sensitive maize genotypes. A subset of four nested association mapping panel parents were selected based on visual leaf scoring for ROS staining assays after submergence. Hydrogen peroxide was detected using stain 3,3′-diaminobenzidine tetrahydrochloride (DAB), while superoxide was detected using nitroblue tetrazolium (NBT). Both assays were performed after 96 h of submergence. In B97, the third leaf was barely emerged and not used for the assay. (b-d) Real-time quantitative PCR of ROS genes. ROS genes were selected based on the study by Gadjev, Vanderauwera and Gechev [ 30 ] and showed a significant induction in response to at least two stresses that induce ROS formation in Arabidopsis. (b.) ALTERNATIVE OXIDASE 1a ( AOX1a ); (c.) WRKY6 (d.) CYTOCHROME P81 D8 ( CYP81D8 ). All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Article Snippet: cDNA synthesis and quantitative PCR First strand cDNA synthesis was performed using SuperScript VILO for real-time quantitative PCR (qPCR) (Invitrogen Corp., Carlsbad, CA, USA).

    Techniques: Expressing, Marker, Staining, Real-time Polymerase Chain Reaction

    PCR and qPCR validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.

    Journal: Frontiers in Plant Science

    Article Title: Mining for Candidate Genes in an Introgression Line by Using RNA Sequencing: The Anthocyanin Overaccumulation Phenotype in Brassica

    doi: 10.3389/fpls.2016.01245

    Figure Lengend Snippet: PCR and qPCR validation of candidate transcripts. (A) PCR amplification (all 36 cycles) of candidate genes from different lines. PDS was used as a positive control. (B) Relative expression levels of green and purple B. juncea ( eEF1B α 2 was used as an internal control). (C) Relative gene expression levels under cold and high light treatment ( eEF1B α 2 was used as an internal control). ** Means statistically significant.

    Article Snippet: Real-time quantitative PCR (qPCR) was performed in a MX3000P qPCR system (Agilent).

    Techniques: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Amplification, Positive Control, Expressing

    Rescue of stress granule formation by Staufen1 down-regulation in DM1 myoblasts. (A) Human DM1 primary myoblasts (GM03987, 500 CTG; and GM03132, 1700 CTG) were infected with a control lentivirus or a specific Staufen1 shRNA. Three days after transductions, cells were treated with 0.5 mM arsenite for 45 min. Immunofluorescence was performed with DDX3 antibodies to visualize SGs. (B) Wild-type primary myoblasts (GM01653 and GM03377) were infected with a control lentivirus expressing GFP or a lentivirus overexpressing Staufen1. Cells were then processed as in A. DAPI was used to stain nuclei. The same exposure conditions were used to ensure quantitative analyses. Scale bars, 20 μm. (C, D) Mean number of SGs per cell. A total of 40 random cells per condition were analyzed. Student’s t tests were used, and asterisks indicate significance (* p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001). (E) Top, relative quantification of Staufen1 mRNA levels as determined by RT-PCR showing up-regulation or knockdown of Staufen1 after lentivirus infections. GAPDH was used to show equal loading. Bottom, IR alternative splicing profiles were assessed by RT-PCR. Numbers show the percentage of IR exon 11 inclusion.

    Journal: Molecular Biology of the Cell

    Article Title: Staufen1 impairs stress granule formation in skeletal muscle cells from myotonic dystrophy type 1 patients

    doi: 10.1091/mbc.E15-06-0356

    Figure Lengend Snippet: Rescue of stress granule formation by Staufen1 down-regulation in DM1 myoblasts. (A) Human DM1 primary myoblasts (GM03987, 500 CTG; and GM03132, 1700 CTG) were infected with a control lentivirus or a specific Staufen1 shRNA. Three days after transductions, cells were treated with 0.5 mM arsenite for 45 min. Immunofluorescence was performed with DDX3 antibodies to visualize SGs. (B) Wild-type primary myoblasts (GM01653 and GM03377) were infected with a control lentivirus expressing GFP or a lentivirus overexpressing Staufen1. Cells were then processed as in A. DAPI was used to stain nuclei. The same exposure conditions were used to ensure quantitative analyses. Scale bars, 20 μm. (C, D) Mean number of SGs per cell. A total of 40 random cells per condition were analyzed. Student’s t tests were used, and asterisks indicate significance (* p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001). (E) Top, relative quantification of Staufen1 mRNA levels as determined by RT-PCR showing up-regulation or knockdown of Staufen1 after lentivirus infections. GAPDH was used to show equal loading. Bottom, IR alternative splicing profiles were assessed by RT-PCR. Numbers show the percentage of IR exon 11 inclusion.

    Article Snippet: A 1-μg amount of RNA was DNase-treated (Ambion/ThermoFisher Scientific, Ottawa, Canada), and cDNAs were synthesized using MuLV Reverse Transcriptase (Applied Biosystems/ThermoFisher Scientific). mRNA expression was evaluated by real-time quantitative PCR (MX3005P; Stratagene/Agilent Technologies, Santa Clara, CA) using the QuantiTect SYBR Green PCR Kit (Qiagen, Toronto, Canada) according to the manufacturer’s instructions.

    Techniques: CTG Assay, Infection, shRNA, Immunofluorescence, Expressing, Staining, Reverse Transcription Polymerase Chain Reaction

    MyoD-conversion induces DMPK expression and aggregation in DM1 cells. (A) Relative quantification of DMPK mRNA levels in fibroblasts and MyoD-converted myoblasts as determined by qRT-PCR. Three independent experiments. (B) FISH of control human DM1 primary fibroblasts and MyoD-converted myoblasts (GM03132, 1700 CTG) using a Cy3-(CAG)10 probe. (C) Quantifications of FISH experiments. From 40 to 57 random cells/condition were analyzed. (D) Top, Western blots showing Staufen1 and CUGBP1 protein levels in wild-type and DM1 fibroblasts and MyoD-converted myoblasts. β-Actin was used as a loading control. Bottom, relative quantification of MyoD mRNA levels as determined by RT-PCR. GAPDH was used to show equal loading. (E) Quantifications of Staufen1 and CUGBP1 levels normalized to GAPDH. Three or four independent experiments. Student’s t tests were used, and asterisks indicate significance (* p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001).

    Journal: Molecular Biology of the Cell

    Article Title: Staufen1 impairs stress granule formation in skeletal muscle cells from myotonic dystrophy type 1 patients

    doi: 10.1091/mbc.E15-06-0356

    Figure Lengend Snippet: MyoD-conversion induces DMPK expression and aggregation in DM1 cells. (A) Relative quantification of DMPK mRNA levels in fibroblasts and MyoD-converted myoblasts as determined by qRT-PCR. Three independent experiments. (B) FISH of control human DM1 primary fibroblasts and MyoD-converted myoblasts (GM03132, 1700 CTG) using a Cy3-(CAG)10 probe. (C) Quantifications of FISH experiments. From 40 to 57 random cells/condition were analyzed. (D) Top, Western blots showing Staufen1 and CUGBP1 protein levels in wild-type and DM1 fibroblasts and MyoD-converted myoblasts. β-Actin was used as a loading control. Bottom, relative quantification of MyoD mRNA levels as determined by RT-PCR. GAPDH was used to show equal loading. (E) Quantifications of Staufen1 and CUGBP1 levels normalized to GAPDH. Three or four independent experiments. Student’s t tests were used, and asterisks indicate significance (* p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001).

    Article Snippet: A 1-μg amount of RNA was DNase-treated (Ambion/ThermoFisher Scientific, Ottawa, Canada), and cDNAs were synthesized using MuLV Reverse Transcriptase (Applied Biosystems/ThermoFisher Scientific). mRNA expression was evaluated by real-time quantitative PCR (MX3005P; Stratagene/Agilent Technologies, Santa Clara, CA) using the QuantiTect SYBR Green PCR Kit (Qiagen, Toronto, Canada) according to the manufacturer’s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Fluorescence In Situ Hybridization, CTG Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction

    Generation of a conditional human JAK2 V617F knock-in allele (A) Diagram showing the endogenous mouse Jak2 allele, the targeting vector, the knock-in allele resulting from homologous recombination, and the activated recombined allele after excision of the PGKNeo-poly (A) cassette. (B) PCR showing high levels of recombination in BM, stem cells, and progenitors and relatively lower levels in peripheral blood, spleen, and thymus. PCR was performed using P1 + P2 + P4 (A) on genomic DNA from peripheral blood (PB), BM, LSK, total progenitors (Prog), LT-HSC (lineage − Sca-1 + c-Kit + CD34 − ), ST-HSC (lineage − Sca-1 + c-Kit + CD34 + ), myeloid progenitors (lineage − Sca-l − c-Kit + CD34 + CD71 − ), and erythroid progenitors (lineage − Sca-l − c-Kit + CD34 − CD71 + ), thymus (Thy), and spleen cells (Sp). Serial dilutions were made by mixing the corresponding amount of genomic DNA from JAK2 F/+ and JAK2 R/+ ES cells. Top (P1 + P2) and bottom (P1 + P4) bands represent the recombined and floxed allele, respectively. (C) PCR (performed as in panel B) showing that the proportion of recombined allele in peripheral blood samples from JAK2 V617F mice increases with time. Vertical lines have been inserted to indicate removal of gel lanes. (D) Real-time quantitative PCR analysis showing comparable up-regulation of Stat5 and Erk1/2 target genes in erythroid cells (both in fetal and adult erythroid cells) from JAK2 V617F mice compared with results obtained from ET patients. In ET patient samples, individual BFU-E were genotyped, pooled according to genotypes, and transcript levels of target genes in JAK2 V617F mutant and wild-type colonies were compared (far right panel). The fold increase represents the ratio of gene expression in V617F and WT pools with each data point representing an individual patient. Transcript levels of the same target genes were increased by a similar amount in both fetal liver and adult BM BFU-E from JAK2 V617F mice (V617F) compared with littermate controls (WT). BFU-E colonies were derived from 3 JAK2 V617F and 3 wild-type control mice. Individual colonies from JAK2 V617F mice were genotyped to distinguish those carrying the active recombined allele. Colonies were pooled according to the genotype (4-6 colonies/pool). Expression of target genes in a pooled BFU-E sample was calculated relative to the mean of the wild-type pooled samples, which was defined as 1. * P

    Journal: Blood

    Article Title: JAK2 V617F impairs hematopoietic stem cell function in a conditional knock-in mouse model of JAK2 V617F-positive essential thrombocythemia

    doi: 10.1182/blood-2009-12-259747

    Figure Lengend Snippet: Generation of a conditional human JAK2 V617F knock-in allele (A) Diagram showing the endogenous mouse Jak2 allele, the targeting vector, the knock-in allele resulting from homologous recombination, and the activated recombined allele after excision of the PGKNeo-poly (A) cassette. (B) PCR showing high levels of recombination in BM, stem cells, and progenitors and relatively lower levels in peripheral blood, spleen, and thymus. PCR was performed using P1 + P2 + P4 (A) on genomic DNA from peripheral blood (PB), BM, LSK, total progenitors (Prog), LT-HSC (lineage − Sca-1 + c-Kit + CD34 − ), ST-HSC (lineage − Sca-1 + c-Kit + CD34 + ), myeloid progenitors (lineage − Sca-l − c-Kit + CD34 + CD71 − ), and erythroid progenitors (lineage − Sca-l − c-Kit + CD34 − CD71 + ), thymus (Thy), and spleen cells (Sp). Serial dilutions were made by mixing the corresponding amount of genomic DNA from JAK2 F/+ and JAK2 R/+ ES cells. Top (P1 + P2) and bottom (P1 + P4) bands represent the recombined and floxed allele, respectively. (C) PCR (performed as in panel B) showing that the proportion of recombined allele in peripheral blood samples from JAK2 V617F mice increases with time. Vertical lines have been inserted to indicate removal of gel lanes. (D) Real-time quantitative PCR analysis showing comparable up-regulation of Stat5 and Erk1/2 target genes in erythroid cells (both in fetal and adult erythroid cells) from JAK2 V617F mice compared with results obtained from ET patients. In ET patient samples, individual BFU-E were genotyped, pooled according to genotypes, and transcript levels of target genes in JAK2 V617F mutant and wild-type colonies were compared (far right panel). The fold increase represents the ratio of gene expression in V617F and WT pools with each data point representing an individual patient. Transcript levels of the same target genes were increased by a similar amount in both fetal liver and adult BM BFU-E from JAK2 V617F mice (V617F) compared with littermate controls (WT). BFU-E colonies were derived from 3 JAK2 V617F and 3 wild-type control mice. Individual colonies from JAK2 V617F mice were genotyped to distinguish those carrying the active recombined allele. Colonies were pooled according to the genotype (4-6 colonies/pool). Expression of target genes in a pooled BFU-E sample was calculated relative to the mean of the wild-type pooled samples, which was defined as 1. * P

    Article Snippet: Real-time quantitative polymerase chain reaction (PCR) was performed using QPCR SYBR Green (Agilent Technologies) and Mx3000P Real-Time PCR system (Stratagene).

    Techniques: Knock-In, Plasmid Preparation, Homologous Recombination, Polymerase Chain Reaction, Mouse Assay, Real-time Polymerase Chain Reaction, Mutagenesis, Expressing, Derivative Assay

    Validation of the accuracy of RNA-seq data. Note: The validation of the accuracy of RNA-seq data was carried out by comparing of RPKM and the relative expression of five randomly selected genes using qRT-PCR for each tissue and time point.

    Journal: PLoS ONE

    Article Title: Identification of Differentially Expressed Genes in Breast Muscle and Skin Fat of Postnatal Pekin Duck

    doi: 10.1371/journal.pone.0107574

    Figure Lengend Snippet: Validation of the accuracy of RNA-seq data. Note: The validation of the accuracy of RNA-seq data was carried out by comparing of RPKM and the relative expression of five randomly selected genes using qRT-PCR for each tissue and time point.

    Article Snippet: Real time quantitative polymerase chain reaction (qRT-PCR) Real time quantitative PCR (qRT-PCR) was performed using the SYBR PrimeScript RT-PCR Kit (TaKaRa) with SYBR Green dye to validate specific gene transcription, RNA-Seq data and variations in gene expression among individuals.

    Techniques: RNA Sequencing Assay, Expressing, Quantitative RT-PCR

    Validation of genetic variation among individuals. Note: The validation of genetic variation among individuals was performed by comparing of the relative expression of ten randomly selected genes within each tissue type and time point using qRT-PCR.

    Journal: PLoS ONE

    Article Title: Identification of Differentially Expressed Genes in Breast Muscle and Skin Fat of Postnatal Pekin Duck

    doi: 10.1371/journal.pone.0107574

    Figure Lengend Snippet: Validation of genetic variation among individuals. Note: The validation of genetic variation among individuals was performed by comparing of the relative expression of ten randomly selected genes within each tissue type and time point using qRT-PCR.

    Article Snippet: Real time quantitative polymerase chain reaction (qRT-PCR) Real time quantitative PCR (qRT-PCR) was performed using the SYBR PrimeScript RT-PCR Kit (TaKaRa) with SYBR Green dye to validate specific gene transcription, RNA-Seq data and variations in gene expression among individuals.

    Techniques: Expressing, Quantitative RT-PCR

    Validation of expression levels of important genes involved in muscle development and fat deposition. Note: The validation of important genes was performed by comparing of RPKM and relative expression using qRT- PCR for each tissue and time point.

    Journal: PLoS ONE

    Article Title: Identification of Differentially Expressed Genes in Breast Muscle and Skin Fat of Postnatal Pekin Duck

    doi: 10.1371/journal.pone.0107574

    Figure Lengend Snippet: Validation of expression levels of important genes involved in muscle development and fat deposition. Note: The validation of important genes was performed by comparing of RPKM and relative expression using qRT- PCR for each tissue and time point.

    Article Snippet: Real time quantitative polymerase chain reaction (qRT-PCR) Real time quantitative PCR (qRT-PCR) was performed using the SYBR PrimeScript RT-PCR Kit (TaKaRa) with SYBR Green dye to validate specific gene transcription, RNA-Seq data and variations in gene expression among individuals.

    Techniques: Expressing, Quantitative RT-PCR

    Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by qRT-PCR, which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of miRNA-19a on gastrointestinal motility in rats with functional dyspepsia

    doi: 10.3892/etm.2018.6009

    Figure Lengend Snippet: Comparison of difference of miRNA-19a expression between model and normal group. The contents of miRNA-19a in rat gastric tissue of model and normal group are detected by qRT-PCR, which displayed that the relative content of miRNA-19a in gastric tissue of model group is 6.13±0.89, which is 4.56±0.65 in normal tissue, suggesting that miRNA-19a is highly expressed in rats with functional dyspepsia, *P

    Article Snippet: TRIzol, reverse-transcription kit and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) kit (Promega Corp., Madison, WI, USA), miRNA-19a, U6 specific primers (Shanghai Sangon Biological Engineering Co., Ltd., Shanghai, China), miRNA-19a sequence mimics (miRNA-19a mimic) and miRNA-19a unrelated sequence mimics (scramble mimic) (Shanghai Sangon Biological Engineering Co., Ltd.), In vivo -jetPEI® (Polyplus Transfection, Illkirch, France), rat motilin (MTL), vasoactive intestinal peptide (VIP) (Wuhan Huamei Biological Engineering Co., Ltd., Wuhan, China), real-time fluorescent quantitative PCR 7500 system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR, Functional Assay

    The inhibitors 17-AAG, EC154, and LBH589 inhibit HIF-dependent gene transcription in CCRCC cells . The CCRCC cell line UMRC2 was treated for 16 h with inhibitors as in Figure 1. Total mRNA was isolated and HIF-α regulated genes analyzed by qRT-PCR. Values were normalized to GAPDH and are presented relative to control cells, with standard deviation shown. All drug treatments significantly reduced all transcript levels (*) as determined by ANOVA and Student's t- test ( p

    Journal: BMC Cancer

    Article Title: Comparative analysis of novel and conventional Hsp90 inhibitors on HIF activity and angiogenic potential in clear cell renal cell carcinoma: implications for clinical evaluation

    doi: 10.1186/1471-2407-11-520

    Figure Lengend Snippet: The inhibitors 17-AAG, EC154, and LBH589 inhibit HIF-dependent gene transcription in CCRCC cells . The CCRCC cell line UMRC2 was treated for 16 h with inhibitors as in Figure 1. Total mRNA was isolated and HIF-α regulated genes analyzed by qRT-PCR. Values were normalized to GAPDH and are presented relative to control cells, with standard deviation shown. All drug treatments significantly reduced all transcript levels (*) as determined by ANOVA and Student's t- test ( p

    Article Snippet: The ratio of GLUT1, VEGF, LOX1 and OCT-4 to α-tubulin was measured with real-time-quantitative PCR (iQ5 Multicolor Real-Time PCR Detection System, Bio-Rad Laboratories, Hercules, CA) using iQ5 optical system software.

    Techniques: Isolation, Quantitative RT-PCR, Standard Deviation

    Time dependent evaluation of 17-AAG, EC154, and LBH589 upon HIF-dependent gene transcription . The CCRCC cell lines UMRC2 and 786-O were treated as in Figure 2 for the indicated times, total mRNA isolated and HIF-α regulated genes analyzed by qRT-PCR. Values were normalized to GAPDH and are presented relative to values at time zero with standard deviation shown.

    Journal: BMC Cancer

    Article Title: Comparative analysis of novel and conventional Hsp90 inhibitors on HIF activity and angiogenic potential in clear cell renal cell carcinoma: implications for clinical evaluation

    doi: 10.1186/1471-2407-11-520

    Figure Lengend Snippet: Time dependent evaluation of 17-AAG, EC154, and LBH589 upon HIF-dependent gene transcription . The CCRCC cell lines UMRC2 and 786-O were treated as in Figure 2 for the indicated times, total mRNA isolated and HIF-α regulated genes analyzed by qRT-PCR. Values were normalized to GAPDH and are presented relative to values at time zero with standard deviation shown.

    Article Snippet: The ratio of GLUT1, VEGF, LOX1 and OCT-4 to α-tubulin was measured with real-time-quantitative PCR (iQ5 Multicolor Real-Time PCR Detection System, Bio-Rad Laboratories, Hercules, CA) using iQ5 optical system software.

    Techniques: Isolation, Quantitative RT-PCR, Standard Deviation

    Effect of labeling on chondrogenic differentiation of ASC. (A) Unlabeled, (B) BNF starch- and (C, E, F) nanomag-D-spio-labeled cells were chondrogenically stimulated for 21 d. Pellets were analyzed using Heidenhain's AZAN trichrome staining showing a reduced collagen type II-positive extracellular matrix (light blue) due to nanoparticle labeling. BNF-labeled cells (25 and 50 µg Fe/ml) failed to generate compact pellets (not shown) (Axio Imager M2, Carl Zeiss Microscopy GmbH, Jena, Germany; scale bars = 100 µm). (D) Collagen type II was analyzed using Real-Time quantitative PCR revealing a diminished mRNA expression of collagen type II due to nanoparticle labeling (2∧ΔΔCT ± %CV; n = 2, normalized to β-actin).

    Journal: PLoS ONE

    Article Title: Comparative In Vitro Study on Magnetic Iron Oxide Nanoparticles for MRI Tracking of Adipose Tissue-Derived Progenitor Cells

    doi: 10.1371/journal.pone.0108055

    Figure Lengend Snippet: Effect of labeling on chondrogenic differentiation of ASC. (A) Unlabeled, (B) BNF starch- and (C, E, F) nanomag-D-spio-labeled cells were chondrogenically stimulated for 21 d. Pellets were analyzed using Heidenhain's AZAN trichrome staining showing a reduced collagen type II-positive extracellular matrix (light blue) due to nanoparticle labeling. BNF-labeled cells (25 and 50 µg Fe/ml) failed to generate compact pellets (not shown) (Axio Imager M2, Carl Zeiss Microscopy GmbH, Jena, Germany; scale bars = 100 µm). (D) Collagen type II was analyzed using Real-Time quantitative PCR revealing a diminished mRNA expression of collagen type II due to nanoparticle labeling (2∧ΔΔCT ± %CV; n = 2, normalized to β-actin).

    Article Snippet: RNA was isolated with NucleoSpin RNA (MACHEREY-NAGEL, Düren, Germany) according to the manufacturer's instructions and the collagen type II expression was analyzed with Real-Time quantitative PCR (qTower 2.0 and qPCR software 1.0, Analytik Jena, Jena, Germany).

    Techniques: Labeling, Staining, Microscopy, Real-time Polymerase Chain Reaction, Expressing

    Magnitude of oval cell response quantified by real time PCR analysis of AFP mRNA, relative to normal liver tissue expression and normalized to beta actin (A), 9AP – animals fed normal rat chow and subjected to 2AAF/PH protocol; 9DAP – animals maintained on the L-cysteine diet associated to 2AAF/PH treatment. GAPDH was used as a loading control (B).

    Journal: Laboratory investigation; a journal of technical methods and pathology

    Article Title: Hepatic stellate cells' involvement in progenitor mediated liver regeneration

    doi: 10.1038/labinvest.2010.88

    Figure Lengend Snippet: Magnitude of oval cell response quantified by real time PCR analysis of AFP mRNA, relative to normal liver tissue expression and normalized to beta actin (A), 9AP – animals fed normal rat chow and subjected to 2AAF/PH protocol; 9DAP – animals maintained on the L-cysteine diet associated to 2AAF/PH treatment. GAPDH was used as a loading control (B).

    Article Snippet: Real time quantitative PCR The mRNA levels were assessed by 2-step quantitative real-time PCR reaction, using a DNA Engine Opticon 2 Continuously Fluorescence Detector (MJ Research Inc Waltham MA).

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    The GM-CSF/IFN-α/VACCINE promoted the clearance of HBcAg-positive hepatocytes ( A ) 14 days after the fourth immunization, liver sections were stained for HBcAg (brown staining) by IHC. Scale bar represents 50 μm. ( B ) 14 days after the fourth immunization, the HBV DNA levels in liver were analyzed by q-PCR. ( C – D ) 14 days after the fourth immunization, serum ALT was measured and the liver-infiltrating lymphocytes were stained by H E. Scale bar represents 50 μm. Symbols represent mean ± SEM. * P

    Journal: Oncotarget

    Article Title: Clearance of HBeAg and HBsAg of HBV in mice model by a recombinant HBV vaccine combined with GM-CSF and IFN-α as an effective therapeutic vaccine adjuvant

    doi: 10.18632/oncotarget.25789

    Figure Lengend Snippet: The GM-CSF/IFN-α/VACCINE promoted the clearance of HBcAg-positive hepatocytes ( A ) 14 days after the fourth immunization, liver sections were stained for HBcAg (brown staining) by IHC. Scale bar represents 50 μm. ( B ) 14 days after the fourth immunization, the HBV DNA levels in liver were analyzed by q-PCR. ( C – D ) 14 days after the fourth immunization, serum ALT was measured and the liver-infiltrating lymphocytes were stained by H E. Scale bar represents 50 μm. Symbols represent mean ± SEM. * P

    Article Snippet: HBV DNA quantitation Serum HBV DNA was determined by real-time quantitative PCR with a kit from Kehua Bio-engineering Co., Ltd (Shanghai, China).

    Techniques: Staining, Immunohistochemistry, Polymerase Chain Reaction