semi-quantitative rt-pcr Search Results


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  • 99
    Qiagen onestep rt pcr kit
    PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). <t>50ng</t> total RNA was used per reaction in all cases for one-step <t>qRT-PCR</t> with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcript analyzed is PhAAE (mean±SE; n =3).
    Onestep Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 11339 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semi quantitative rt pcr taqman microrna assays
    PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). <t>50ng</t> total RNA was used per reaction in all cases for one-step <t>qRT-PCR</t> with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcript analyzed is PhAAE (mean±SE; n =3).
    Semi Quantitative Rt Pcr Taqman Microrna Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcription polymerase chain reaction rt pcr semi quantitative rt pcr
    PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). <t>50ng</t> total RNA was used per reaction in all cases for one-step <t>qRT-PCR</t> with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcript analyzed is PhAAE (mean±SE; n =3).
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Semi Quantitative Rt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega semi quantitative rt pcr
    Key cardiovascular gene expression gradient in a chick embryo. ( A ) Super-position of the different cardiovascular factors on a single embryo based on in situ hybridization and semi <t>qRT-PCR</t> analysis. ( B ) Semi qRT-PCR analysis of cardiovascular genes in a St.8 - embryo. The embryo is divided to four sections that are manually dissected and subjected to <t>RNA</t> analysis. Representative results of three different biological repeats. lpm – lateral plate mesoderm; np – neural plate; cc – cardiac crescent; ee – extraembryonic tissue. DOI: http://dx.doi.org/10.7554/eLife.20994.019
    Semi Quantitative Rt Pcr, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 374 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad semi quantitative rt pcr
    Impacts of genetic manipulation of IAHs on plant sensitivity to exogenous IAA conjugates. (A–C) Photographs and root length measurements of 9-day-old WT, ILL6-OE (line 5), IAR3-OE (line 42), ILR1-OE , ill6-2 , iar3-5 , and ilr1-5 plants grown on MS medium supplemented or not with 5 μM IAA, 50 μM IAA-Ala, or 30 μM IAA-Leu. Data show the mean and SD ( n > 18). (D and E) <t>qRT–PCR</t> analysis of IAA5 and GH3.1 expression in leaves of 4-week-old WT and ILL6-OE plants sprayed (2h) with the mock, 5 μM IAA, or 50 μM IAA-Ala solutions. Relative abundance compared with ACTIN8 is displayed. Error bars indicate the SD of three biological replicates. Asterisks denote a significant difference compared with the WT at P
    Semi Quantitative Rt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 311 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa semi quantitative reverse transcription polymerase chain reaction rt pcr
    Five genes with AS events were affected by GA 3 . a - c . Wiggle track for three candidate genes, which have differential isoforms after GA3 treated. Gene structures in blue originated from Pacbio SMRT dataset, while green were generated by Cufflinks assemble of <t>RNA-Seq.</t> “P” indicates the AS events predicted with Pacbio SMRT data, and “C” represented events from RNA-Seq data. d . AS events from five genes were validated by <t>RT-PCR</t> gel blot with UBQ as the positive control. “+” indicated GA treatment, and “-” represented non-treatment. e . Validation of AS events by qRT-PCR
    Semi Quantitative Reverse Transcription Polymerase Chain Reaction Rt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore semi quantitative rt pcr
    Five genes with AS events were affected by GA 3 . a - c . Wiggle track for three candidate genes, which have differential isoforms after GA3 treated. Gene structures in blue originated from Pacbio SMRT dataset, while green were generated by Cufflinks assemble of <t>RNA-Seq.</t> “P” indicates the AS events predicted with Pacbio SMRT data, and “C” represented events from RNA-Seq data. d . AS events from five genes were validated by <t>RT-PCR</t> gel blot with UBQ as the positive control. “+” indicated GA treatment, and “-” represented non-treatment. e . Validation of AS events by qRT-PCR
    Semi Quantitative Rt Pcr, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semi quantitative rt pcr trizol reagent
    Five genes with AS events were affected by GA 3 . a - c . Wiggle track for three candidate genes, which have differential isoforms after GA3 treated. Gene structures in blue originated from Pacbio SMRT dataset, while green were generated by Cufflinks assemble of <t>RNA-Seq.</t> “P” indicates the AS events predicted with Pacbio SMRT data, and “C” represented events from RNA-Seq data. d . AS events from five genes were validated by <t>RT-PCR</t> gel blot with UBQ as the positive control. “+” indicated GA treatment, and “-” represented non-treatment. e . Validation of AS events by qRT-PCR
    Semi Quantitative Rt Pcr Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semi quantitative one step rt pcr
    Transcriptional analysis of genes upstream and downstream from five clonal mutants. Genomic DNA-free total RNA from wild-type E . chaffeensis (lane 1) and clonal mutants (lane 2) was assessed for the genes 5’ and 3’ to insertion sites by semi quantitative <t>RT-PCR.</t> Panels 1A-5A have the cartoons depicting the insertions and their orientations within the genome and the genes near the insertions. The annotated/predicted gene names near the insertions were provided in each sub-panel. Panels 1B-5B had RT-PCR products resolved on agarose gels for each of the respective genes as in panels 1A-5A. No template controls (lane 3) and wild type genomic DNA template reactions (lane 4) were included to serve as negative and positive controls for each reaction, respectively. M represents 1 kb plus DNA ladder (Life Technologies. <t>Carlsbad,</t> CA). Molecular sizes of the resolved DNA ladder shown in the Figs are the same for each sub-panels and were identified in the left image of panel 1B. Predicted amplicon for each gene was listed in Table 1 . The primer binding locations relative to whole genome data were also provided in Table 1 .
    Semi Quantitative One Step Rt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semi quantitative rt pcr method
    PEA attenuates Aβ-induced astrocyte activation. Aβ-challenged (1 μg/ml) cells were treated with PEA (10 −7 M) in the presence of the selective PPAR-γ antagonist (GW9662, 9 nM) or the selective PPAR-α antagonist (MK886, 3 μM). GFAP <t>mRNA</t> was evaluated 12 hrs following Aβ challenge; GFAP and S100B protein expression was evaluated after 24 hrs of treatments by Western blot and immunofluorescence analyses. S100B release in the cellular milieu was determined 24 hrs after Aβ challenge by ELISA assay. (A) Results of GFAP and S100B Western blot analysis and densitometric analysis of corresponding bands. β-actin was used as loading control. (B) Results of GFAP <t>RT-PCR</t> amplification and densitometric analysis of corresponding bands. GAPDH was used as standard control. (C) Measurement of S100B release by ELISA assay. (D) Analysis of GFAP and S100B protein expression by immunofluorescence (magnification 10 × ). Results are the mean ± S.E.M. of n = 4 separate experiments. *** P
    Semi Quantitative Rt Pcr Method, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems semi quantitative rt pcr
    PEA attenuates Aβ-induced astrocyte activation. Aβ-challenged (1 μg/ml) cells were treated with PEA (10 −7 M) in the presence of the selective PPAR-γ antagonist (GW9662, 9 nM) or the selective PPAR-α antagonist (MK886, 3 μM). GFAP <t>mRNA</t> was evaluated 12 hrs following Aβ challenge; GFAP and S100B protein expression was evaluated after 24 hrs of treatments by Western blot and immunofluorescence analyses. S100B release in the cellular milieu was determined 24 hrs after Aβ challenge by ELISA assay. (A) Results of GFAP and S100B Western blot analysis and densitometric analysis of corresponding bands. β-actin was used as loading control. (B) Results of GFAP <t>RT-PCR</t> amplification and densitometric analysis of corresponding bands. GAPDH was used as standard control. (C) Measurement of S100B release by ELISA assay. (D) Analysis of GFAP and S100B protein expression by immunofluorescence (magnification 10 × ). Results are the mean ± S.E.M. of n = 4 separate experiments. *** P
    Semi Quantitative Rt Pcr, supplied by R&D Systems, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim semi quantitative rt pcr
    Fig. 5. Quantitation of the hypertrophic phenotype of AdMEK1-infected cardiomyocytes. ( A ) Cell surface area was measured in α-actinin-stained cardiomyocyte cultures using confocal microscopy and digitized imaging (24 h after infection). Cultures were left in serum-free medium with no stimulation or were infected with Adβgal or AdMEK1, or were treated with U0126. ( B ) Cardiomyocytes were also stained with an <t>ANF-specific</t> antibody to quantify the percentage of cells expressing ANF. The data in (A) and (B) were obtained in two independent experiments. ( C ) Total ANF mRNA levels were also quantified by <t>RT–PCR,</t> showing similar induction of ANF mRNA between 1% FBS and AdMEK1 infection. ( D ) Protein synthesis rates were monitored by the incorporation of [ 3 H]leucine in Adβgal-, AdMEK1- and 1% FBS-stimulated cardiomyocytes. * P
    Semi Quantitative Rt Pcr, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MJ Research semi quantitative rt pcr
    <t>RT–PCR</t> of <t>AQP4</t> of striatum (a), and of cortex (b) of different set of experimental rats. M denotes marker i.e. DNA ladder. C denotes negative control. S denotes sham groups, S+D denote sham treated with drug, V denotes the vehicle and V+D denote the vehicle treated with drug. Histograms represent cumulative data expressed as mean ± SEM obtained from three different sets of experiments conducted for striatum and cortex. ( a vs sham b vs vehicle) ( P ≤0.05).
    Semi Quantitative Rt Pcr, supplied by MJ Research, used in various techniques. Bioz Stars score: 98/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioneer Corporation semi quantitative rt pcr
    <t>RT–PCR</t> of <t>AQP4</t> of striatum (a), and of cortex (b) of different set of experimental rats. M denotes marker i.e. DNA ladder. C denotes negative control. S denotes sham groups, S+D denote sham treated with drug, V denotes the vehicle and V+D denote the vehicle treated with drug. Histograms represent cumulative data expressed as mean ± SEM obtained from three different sets of experiments conducted for striatum and cortex. ( a vs sham b vs vehicle) ( P ≤0.05).
    Semi Quantitative Rt Pcr, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 99/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GraphPad Prism Inc semi quantitative rt pcr
    <t>RT–PCR</t> of <t>AQP4</t> of striatum (a), and of cortex (b) of different set of experimental rats. M denotes marker i.e. DNA ladder. C denotes negative control. S denotes sham groups, S+D denote sham treated with drug, V denotes the vehicle and V+D denote the vehicle treated with drug. Histograms represent cumulative data expressed as mean ± SEM obtained from three different sets of experiments conducted for striatum and cortex. ( a vs sham b vs vehicle) ( P ≤0.05).
    Semi Quantitative Rt Pcr, supplied by GraphPad Prism Inc, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies semi quantitative rt pcr rt pcr
    (A) <t>qRT-PCR</t> amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no <t>miRNA</t> was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.
    Semi Quantitative Rt Pcr Rt Pcr, supplied by Agilent technologies, 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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    Kodak Corp semi quantitative rt pcr
    (A) <t>qRT-PCR</t> amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no <t>miRNA</t> was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.
    Semi Quantitative Rt Pcr, supplied by Kodak Corp, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semi quantitative rt pcr reagent
    (A) <t>qRT-PCR</t> amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no <t>miRNA</t> was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.
    Semi Quantitative Rt Pcr Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co semi quantitative rt pcr analysis
    (A) <t>qRT-PCR</t> amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no <t>miRNA</t> was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.
    Semi Quantitative Rt Pcr Analysis, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Altona Diagnostics semi quantitative rt pcr assay
    JIKI trial mortality, according to age and baseline <t>RT-PCR</t> Ct value. Histograms represent mortality percentages. Vertical bars represent 95% confidence intervals. Red bars represent target values. The RT-PCR assay was conducted using the <t>RealStar</t> Filovirus Screen RT-PCR Kit 1.0 (Altona Diagnostics).
    Semi Quantitative Rt Pcr Assay, supplied by Altona Diagnostics, 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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    Illumina Inc semi quantitative rt pcr analysis
    JIKI trial mortality, according to age and baseline <t>RT-PCR</t> Ct value. Histograms represent mortality percentages. Vertical bars represent 95% confidence intervals. Red bars represent target values. The RT-PCR assay was conducted using the <t>RealStar</t> Filovirus Screen RT-PCR Kit 1.0 (Altona Diagnostics).
    Semi Quantitative Rt Pcr Analysis, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 84/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa semi quantitative rt pcr tissue total rnas
    Tissue-restricted expression of ELA in the human. Reverse transcription <t>PCR</t> was conducted on total <t>RNAs</t> extracted from the indicated human tissues, and embryonic (ESCs) and induced (iPSCs) pluripotent stem cells using primers specific for human ELA, APJ and β-actin.
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    Santa Cruz Biotechnology semi quantitative nested rt pcr
    Tissue-restricted expression of ELA in the human. Reverse transcription <t>PCR</t> was conducted on total <t>RNAs</t> extracted from the indicated human tissues, and embryonic (ESCs) and induced (iPSCs) pluripotent stem cells using primers specific for human ELA, APJ and β-actin.
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    Eppendorf AG semi quantitative rt pcr reactions
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
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    Thermo Fisher semi quantitative rt pcr trizol
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
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    Syngene semi quantitative rt pcr results
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
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    Thermo Fisher semi quantitative rt pcr analysis sybr green chemistry
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
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    Millipore semi quantitative rt pcr trizol reagent
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
    Semi Quantitative Rt Pcr Trizol Reagent, supplied by Millipore, 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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    TaKaRa semi quantitative rt pcr analysis trizol
    <t>qRT-PCR</t> analysis of Heat shock <t>protein70</t> gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P
    Semi Quantitative Rt Pcr Analysis Trizol, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vivo NHP infection with B. burgdorferi increases NK-1R expression in the CNS, and such increases are prevented by treatment with the NK-1R antagonist aprepitant. Rhesus macaques were uninfected ( n = 2 animals) or infected intrathecally with B. burgdorferi (Bb, 1 × 10 8 bacteria; n = 8), and infected animals were either untreated ( n = 4) or treated with aprepitant ( n = 4) for 2 or 4 weeks prior to euthanasia. Expression of <t>mRNA</t> encoding NK-1R in frontal cortical tissue samples was determined by <t>RT-PCR</t> ( a ) and relative expression normalized to GAPDH levels was determined by densitometric analysis ( b ). NK-1R protein expression was determined in tissue samples by immunoblot analysis and normalized to β-actin expression ( c ). Data is expressed as the mean ± SD. Asterisk and pound symbols indicate statistically significant difference from uninfected animals and untreated infected animals, respectively ( p
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    Millipore semi quantitative rt pcr total rna isolation
    In vivo NHP infection with B. burgdorferi increases NK-1R expression in the CNS, and such increases are prevented by treatment with the NK-1R antagonist aprepitant. Rhesus macaques were uninfected ( n = 2 animals) or infected intrathecally with B. burgdorferi (Bb, 1 × 10 8 bacteria; n = 8), and infected animals were either untreated ( n = 4) or treated with aprepitant ( n = 4) for 2 or 4 weeks prior to euthanasia. Expression of <t>mRNA</t> encoding NK-1R in frontal cortical tissue samples was determined by <t>RT-PCR</t> ( a ) and relative expression normalized to GAPDH levels was determined by densitometric analysis ( b ). NK-1R protein expression was determined in tissue samples by immunoblot analysis and normalized to β-actin expression ( c ). Data is expressed as the mean ± SD. Asterisk and pound symbols indicate statistically significant difference from uninfected animals and untreated infected animals, respectively ( p
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    Clustering of the differentially expressed proteins and a comparison of the expression patterns for ten proteins and their transcripts. The hierarchical clustering of the abundance volume of the protein spots ( A ) and their relative volume ratios ( B ) at S3 on 2-DE gels are shown. The average changed abundance ratios of the identified protein spots on 2-D DIGE gels are also presented ( C ). The high- or low-abundance protein spots or the induced- or reduced-values for the relative changed ratios were indicated in red or green, respectively. The details for hierarchical clustering of the changed patterns of the differential protein spots on 2-DE and 2-D DIGE are presented in Table S2 and Table S3 , respectively. Then, 10 DEPs with sharp changes were selected to determine the correlations between their gene and protein expression levels using <t>RT-PCR</t> and <t>qRT-PCR.</t> The spot numbers for a specific protein are given on the right side of the heat plot next to the same protein in the heat plot. For each treatment, the expression of each gene is presented as a ratio between the gene value and the corresponding 18SRNA value. The gray dot line in each rectangle graph represents a 1.0 ratio value. The error bars indicate the SD of three independent PCR reactions for three biological replicates. For UDXS and MADH, antibodies were used to determine the protein expression, revealing that the two proteins had similar expression patterns to those on the 2-DE gels ( D ). Abbreviations: AGPS, ADP-glucose pyrophosphorylase small subunit; GLUP, glucan phosphorylase; APX1, ascorbate peroxidase-1; initiation factor eIF5-A; SOD, superoxide dismutase; SAMS, S-adenosylmethionine synthetase; UDXS, UDP-D-apiose/UDP-D-xylose synthase; MADH, malate dehydrogenase; WB, Western blotting.
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    Image Search Results


    PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). 50ng total RNA was used per reaction in all cases for one-step qRT-PCR with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcript analyzed is PhAAE (mean±SE; n =3).

    Journal: Journal of Experimental Botany

    Article Title: A peroxisomally localized acyl-activating enzyme is required for volatile benzenoid formation in a Petunia×hybrida cv. 'Mitchell Diploid' flower

    doi: 10.1093/jxb/ers153

    Figure Lengend Snippet: PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). 50ng total RNA was used per reaction in all cases for one-step qRT-PCR with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcript analyzed is PhAAE (mean±SE; n =3).

    Article Snippet: Semi-quantitative (sq)RT-PCR was performed using a Qiagen One-step RT-PCR kit (Qiagen Co., Valencia, CA, USA) with 50ng total RNA.

    Techniques: Quantitative RT-PCR, Isolation

    PhAAE transcript accumulation analysis (one-step qRT-PCR). Spatial analysis used root, stem, stigma, anther, leaf, petal tube, petal limb, and sepal tissues of MD harvested at 16.00h (A). Floral developmental analysis used MD flowers from 11 sequential stages collected on one day at 16.00h (B). Ethylene treatment (2 µl l –1 analysis used excised MD and 44 568 whole flowers treated for 0, 1, 2, 4, and 8h (C, D). 50ng total RNA was used per reaction in all cases. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references (mean±SE; n =3).

    Journal: Journal of Experimental Botany

    Article Title: A peroxisomally localized acyl-activating enzyme is required for volatile benzenoid formation in a Petunia×hybrida cv. 'Mitchell Diploid' flower

    doi: 10.1093/jxb/ers153

    Figure Lengend Snippet: PhAAE transcript accumulation analysis (one-step qRT-PCR). Spatial analysis used root, stem, stigma, anther, leaf, petal tube, petal limb, and sepal tissues of MD harvested at 16.00h (A). Floral developmental analysis used MD flowers from 11 sequential stages collected on one day at 16.00h (B). Ethylene treatment (2 µl l –1 analysis used excised MD and 44 568 whole flowers treated for 0, 1, 2, 4, and 8h (C, D). 50ng total RNA was used per reaction in all cases. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references (mean±SE; n =3).

    Article Snippet: Semi-quantitative (sq)RT-PCR was performed using a Qiagen One-step RT-PCR kit (Qiagen Co., Valencia, CA, USA) with 50ng total RNA.

    Techniques: Quantitative RT-PCR

    PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). 50ng total RNA was used per reaction in all cases for one-step qRT-PCR with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcripts analyzed are PhBSMT , PhBPBT , PhCFAT , PhIGS1 , PhPAAS , PhKAT1 , PhCM1 , PhPAL1 , PhPAL2 , PhODO1 , PhC4H1 , PhC4H2 , and PhMYB4 (mean±SE; n =3).

    Journal: Journal of Experimental Botany

    Article Title: A peroxisomally localized acyl-activating enzyme is required for volatile benzenoid formation in a Petunia×hybrida cv. 'Mitchell Diploid' flower

    doi: 10.1093/jxb/ers153

    Figure Lengend Snippet: PhAAE comparative transcript accumulation analysis between MD and two independent, homozygous T 2 ir-PhAAE lines (15.15 and 24.8). 50ng total RNA was used per reaction in all cases for one-step qRT-PCR with RNA isolated from stage 8 flowers at 16.00h. Histograms are representative of multiple experiments and multiple biological replicates, and analyzed by the ∆∆Ct method with PhFBP1 and Ph18S as the internal references. The individual petunia transcripts analyzed are PhBSMT , PhBPBT , PhCFAT , PhIGS1 , PhPAAS , PhKAT1 , PhCM1 , PhPAL1 , PhPAL2 , PhODO1 , PhC4H1 , PhC4H2 , and PhMYB4 (mean±SE; n =3).

    Article Snippet: Semi-quantitative (sq)RT-PCR was performed using a Qiagen One-step RT-PCR kit (Qiagen Co., Valencia, CA, USA) with 50ng total RNA.

    Techniques: Quantitative RT-PCR, Isolation

    Key cardiovascular gene expression gradient in a chick embryo. ( A ) Super-position of the different cardiovascular factors on a single embryo based on in situ hybridization and semi qRT-PCR analysis. ( B ) Semi qRT-PCR analysis of cardiovascular genes in a St.8 - embryo. The embryo is divided to four sections that are manually dissected and subjected to RNA analysis. Representative results of three different biological repeats. lpm – lateral plate mesoderm; np – neural plate; cc – cardiac crescent; ee – extraembryonic tissue. DOI: http://dx.doi.org/10.7554/eLife.20994.019

    Journal: eLife

    Article Title: Nkx2.5 marks angioblasts that contribute to hemogenic endothelium of the endocardium and dorsal aorta

    doi: 10.7554/eLife.20994

    Figure Lengend Snippet: Key cardiovascular gene expression gradient in a chick embryo. ( A ) Super-position of the different cardiovascular factors on a single embryo based on in situ hybridization and semi qRT-PCR analysis. ( B ) Semi qRT-PCR analysis of cardiovascular genes in a St.8 - embryo. The embryo is divided to four sections that are manually dissected and subjected to RNA analysis. Representative results of three different biological repeats. lpm – lateral plate mesoderm; np – neural plate; cc – cardiac crescent; ee – extraembryonic tissue. DOI: http://dx.doi.org/10.7554/eLife.20994.019

    Article Snippet: The RNA analysis was performed using semi-quantitative RT-PCR with Green Master Mix (Promega, Madison, WI USA).

    Techniques: Expressing, In Situ Hybridization, Quantitative RT-PCR

    Impacts of genetic manipulation of IAHs on plant sensitivity to exogenous IAA conjugates. (A–C) Photographs and root length measurements of 9-day-old WT, ILL6-OE (line 5), IAR3-OE (line 42), ILR1-OE , ill6-2 , iar3-5 , and ilr1-5 plants grown on MS medium supplemented or not with 5 μM IAA, 50 μM IAA-Ala, or 30 μM IAA-Leu. Data show the mean and SD ( n > 18). (D and E) qRT–PCR analysis of IAA5 and GH3.1 expression in leaves of 4-week-old WT and ILL6-OE plants sprayed (2h) with the mock, 5 μM IAA, or 50 μM IAA-Ala solutions. Relative abundance compared with ACTIN8 is displayed. Error bars indicate the SD of three biological replicates. Asterisks denote a significant difference compared with the WT at P

    Journal: Journal of Experimental Botany

    Article Title: Hormone crosstalk in wound stress response: wound-inducible amidohydrolases can simultaneously regulate jasmonate and auxin homeostasis in Arabidopsis thaliana

    doi: 10.1093/jxb/erv521

    Figure Lengend Snippet: Impacts of genetic manipulation of IAHs on plant sensitivity to exogenous IAA conjugates. (A–C) Photographs and root length measurements of 9-day-old WT, ILL6-OE (line 5), IAR3-OE (line 42), ILR1-OE , ill6-2 , iar3-5 , and ilr1-5 plants grown on MS medium supplemented or not with 5 μM IAA, 50 μM IAA-Ala, or 30 μM IAA-Leu. Data show the mean and SD ( n > 18). (D and E) qRT–PCR analysis of IAA5 and GH3.1 expression in leaves of 4-week-old WT and ILL6-OE plants sprayed (2h) with the mock, 5 μM IAA, or 50 μM IAA-Ala solutions. Relative abundance compared with ACTIN8 is displayed. Error bars indicate the SD of three biological replicates. Asterisks denote a significant difference compared with the WT at P

    Article Snippet: Phusion Taq polymerase (Thermo Scientific) and Taq-Pro Red Complete (Denville) were used as DNA polymerases for cloning purposes and for semi-quantitative RT–PCR experiments, respectively. qRT–PCR was performed on a CFX96 Touch™ real-time PCR detection system (Bio-Rad) using SsoFast™ EvaGreen® Supermix (Bio-Rad) as per the manufacturer’s instructions.

    Techniques: Mass Spectrometry, Quantitative RT-PCR, Expressing

    Overexpression of amidohydrolases impacts JA and IAA marker gene expression. (A and B) qRT–PCR analysis of JAZ8 expression in unwounded and wounded (2h) leaves of the WT, ILL6-OE (line 5), IAR3-OE (line 42), and ILR1-OE . Fold change relative to the unwounded WT transcript level is displayed. (C and D) IAA5 expression in 9-day-old seedlings grown on MS medium containing the mock treatment, 50 μM IAA-Ala, or 30 μM IAA-Leu. Expression levels relative to ACTIN8 are displayed. Error bars denote the SD of three biological replicates. Asterisks indicate a significant difference at P

    Journal: Journal of Experimental Botany

    Article Title: Hormone crosstalk in wound stress response: wound-inducible amidohydrolases can simultaneously regulate jasmonate and auxin homeostasis in Arabidopsis thaliana

    doi: 10.1093/jxb/erv521

    Figure Lengend Snippet: Overexpression of amidohydrolases impacts JA and IAA marker gene expression. (A and B) qRT–PCR analysis of JAZ8 expression in unwounded and wounded (2h) leaves of the WT, ILL6-OE (line 5), IAR3-OE (line 42), and ILR1-OE . Fold change relative to the unwounded WT transcript level is displayed. (C and D) IAA5 expression in 9-day-old seedlings grown on MS medium containing the mock treatment, 50 μM IAA-Ala, or 30 μM IAA-Leu. Expression levels relative to ACTIN8 are displayed. Error bars denote the SD of three biological replicates. Asterisks indicate a significant difference at P

    Article Snippet: Phusion Taq polymerase (Thermo Scientific) and Taq-Pro Red Complete (Denville) were used as DNA polymerases for cloning purposes and for semi-quantitative RT–PCR experiments, respectively. qRT–PCR was performed on a CFX96 Touch™ real-time PCR detection system (Bio-Rad) using SsoFast™ EvaGreen® Supermix (Bio-Rad) as per the manufacturer’s instructions.

    Techniques: Over Expression, Marker, Expressing, Quantitative RT-PCR, Mass Spectrometry

    SB203580 enhances G-CSF expression in LPS-treated macrophages. RAW264.7 cells ( a and b ) and BMDMs ( c and d ) were incubated with DMSO (vehicle) or 10 μM SB203580 for 30 min, then 100 ng/ml of LPS was added and incubation continued for 0 to 16 h, then G-CSF protein levels in the medium were measured by ELISA ( a and c ) and G-CSF and IL-1β mRNA levels were determined by RT-PCR and analyzed by gel electrophoresis ( b and d ). ( e ) RAW264.7 cells were incubated with DMSO or 1, 5, or 10 μM SB203580 for 30 min, then LPS (100 ng/ml) was added and incubation continued for 6 h, then G-CSF and IL-1β mRNA levels were determined by RT-PCR ( upper panel ). Levels of G-CSF mRNA were determined by RT-qPCR ( lower panel ), normalized to GAPDH mRNA levels, and expressed relative to the value in the cells treated only with LPS, set as 1. ( f ) THP-1 macrophages were pretreated with DMSO or 10 μM SB203580 for 30 min, then LPS or PBS was added and incubation continued for 0 to 6 h. Total RNA was then isolated and levels of G-CSF and GAPDH (internal control) mRNA determined by RT-PCR and analyzed by gel electrophoresis. The results in ( a ) and ( e ) are the mean ± SD for three independent experiments. * p

    Journal: Journal of Biomedical Science

    Article Title: SB203580 increases G-CSF production via a stem-loop destabilizing element in the 3’ untranslated region in macrophages independently of its effect on p38 MAPK activity

    doi: 10.1186/s12929-016-0221-z

    Figure Lengend Snippet: SB203580 enhances G-CSF expression in LPS-treated macrophages. RAW264.7 cells ( a and b ) and BMDMs ( c and d ) were incubated with DMSO (vehicle) or 10 μM SB203580 for 30 min, then 100 ng/ml of LPS was added and incubation continued for 0 to 16 h, then G-CSF protein levels in the medium were measured by ELISA ( a and c ) and G-CSF and IL-1β mRNA levels were determined by RT-PCR and analyzed by gel electrophoresis ( b and d ). ( e ) RAW264.7 cells were incubated with DMSO or 1, 5, or 10 μM SB203580 for 30 min, then LPS (100 ng/ml) was added and incubation continued for 6 h, then G-CSF and IL-1β mRNA levels were determined by RT-PCR ( upper panel ). Levels of G-CSF mRNA were determined by RT-qPCR ( lower panel ), normalized to GAPDH mRNA levels, and expressed relative to the value in the cells treated only with LPS, set as 1. ( f ) THP-1 macrophages were pretreated with DMSO or 10 μM SB203580 for 30 min, then LPS or PBS was added and incubation continued for 0 to 6 h. Total RNA was then isolated and levels of G-CSF and GAPDH (internal control) mRNA determined by RT-PCR and analyzed by gel electrophoresis. The results in ( a ) and ( e ) are the mean ± SD for three independent experiments. * p

    Article Snippet: Levels of G-CSF and GAPDH mRNA were determined either by semi-quantitative PCR (RT-PCR) or by real-time quantitative PCR (RT-qPCR) on an iCycler instrument (BioRad, Hercules, CA, USA) as indicated.

    Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Quantitative RT-PCR, Isolation

    SB203580 induces G-CSF expression in RAW264.7 cells in the absence of LPS stimulation. a RAW264.7 cells were left untreated (lane 1) or were incubated with LPS for 15 to 60 min (lanes 2–4), then phosphorylated p38, total p38, and β-actin levels were analyzed by Western blotting. b RAW264.7 cells were incubated with DMSO or 10 μM SB203580 for 0 to 8 h, then levels of G-CSF and GAPDH (internal control) mRNA were determined by RT-PCR and analyzed by gel electrophoresis ( upper panels ). The G-CSF mRNA levels were normalized to those for GAPDH mRNA and expressed relative to the value in the DMSO-treated cells at 0 h (relative value = 1) ( lower panels ). The results are the mean ± SD for three independent experiments. * p

    Journal: Journal of Biomedical Science

    Article Title: SB203580 increases G-CSF production via a stem-loop destabilizing element in the 3’ untranslated region in macrophages independently of its effect on p38 MAPK activity

    doi: 10.1186/s12929-016-0221-z

    Figure Lengend Snippet: SB203580 induces G-CSF expression in RAW264.7 cells in the absence of LPS stimulation. a RAW264.7 cells were left untreated (lane 1) or were incubated with LPS for 15 to 60 min (lanes 2–4), then phosphorylated p38, total p38, and β-actin levels were analyzed by Western blotting. b RAW264.7 cells were incubated with DMSO or 10 μM SB203580 for 0 to 8 h, then levels of G-CSF and GAPDH (internal control) mRNA were determined by RT-PCR and analyzed by gel electrophoresis ( upper panels ). The G-CSF mRNA levels were normalized to those for GAPDH mRNA and expressed relative to the value in the DMSO-treated cells at 0 h (relative value = 1) ( lower panels ). The results are the mean ± SD for three independent experiments. * p

    Article Snippet: Levels of G-CSF and GAPDH mRNA were determined either by semi-quantitative PCR (RT-PCR) or by real-time quantitative PCR (RT-qPCR) on an iCycler instrument (BioRad, Hercules, CA, USA) as indicated.

    Techniques: Expressing, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Nucleic Acid Electrophoresis

    Five genes with AS events were affected by GA 3 . a - c . Wiggle track for three candidate genes, which have differential isoforms after GA3 treated. Gene structures in blue originated from Pacbio SMRT dataset, while green were generated by Cufflinks assemble of RNA-Seq. “P” indicates the AS events predicted with Pacbio SMRT data, and “C” represented events from RNA-Seq data. d . AS events from five genes were validated by RT-PCR gel blot with UBQ as the positive control. “+” indicated GA treatment, and “-” represented non-treatment. e . Validation of AS events by qRT-PCR

    Journal: BMC Plant Biology

    Article Title: Transcriptome characterization of moso bamboo (Phyllostachys edulis) seedlings in response to exogenous gibberellin applications

    doi: 10.1186/s12870-018-1336-z

    Figure Lengend Snippet: Five genes with AS events were affected by GA 3 . a - c . Wiggle track for three candidate genes, which have differential isoforms after GA3 treated. Gene structures in blue originated from Pacbio SMRT dataset, while green were generated by Cufflinks assemble of RNA-Seq. “P” indicates the AS events predicted with Pacbio SMRT data, and “C” represented events from RNA-Seq data. d . AS events from five genes were validated by RT-PCR gel blot with UBQ as the positive control. “+” indicated GA treatment, and “-” represented non-treatment. e . Validation of AS events by qRT-PCR

    Article Snippet: Experimental validation by semi-quantitative RT-PCR 1 μg RNA was used for cDNA synthesis with PrimeScript™ RT reagent Kit with gDNA Eraser (TaKaRa, Cat#RR047A), and PCR was conducted with Premix Taq (TaKaRa Taq Version 2.0 plus dye,Cat#RR901A) in a total 30 μl reaction volume with UBQ as the positive control.

    Techniques: Generated, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Positive Control, Quantitative RT-PCR

    Expression pattern of BhbZIP60. (A–C) qRT-PCR analysis of the expression of BhbZIP60U and BhbZIP60S under dehydration (A) , 0.5 μg ml -1 TM (B) , and 100 μM ABA, 150 mM NaCl and 20 mM MV for 24 h, and 42°C heat shock for 1 h (C) . Three-month old B. hygrometrica were used for various treatments with 18S rRNA gene used as the internal control. Data were expressed as the mean ± SD of three independent experiments. Different letters indicate P

    Journal: Frontiers in Plant Science

    Article Title: BhbZIP60 from Resurrection Plant Boea hygrometrica Is an mRNA Splicing-Activated Endoplasmic Reticulum Stress Regulator Involved in Drought Tolerance

    doi: 10.3389/fpls.2017.00245

    Figure Lengend Snippet: Expression pattern of BhbZIP60. (A–C) qRT-PCR analysis of the expression of BhbZIP60U and BhbZIP60S under dehydration (A) , 0.5 μg ml -1 TM (B) , and 100 μM ABA, 150 mM NaCl and 20 mM MV for 24 h, and 42°C heat shock for 1 h (C) . Three-month old B. hygrometrica were used for various treatments with 18S rRNA gene used as the internal control. Data were expressed as the mean ± SD of three independent experiments. Different letters indicate P

    Article Snippet: Semi-Quantitative RT-PCR (RT-PCR) and Quantitative Real-Time PCR (qRT-PCR) Analysis Total RNA was extracted using Trizol reagent (TaKaRa).

    Techniques: Expressing, Quantitative RT-PCR

    Complementation experiment of Arabidopsis atbzip60 mutant. Complementation of atbzip60 by BhbZIP60S was demonstrated by the expression of UPR genes detected using qRT-PCR. OE(atbzip60)-1 and OE(atbzip60)-2 represent overexpressing BhbZIP60S in atbzip60 background. 18S rRNA was used as an internal reference gene. Data represent the means ± SD of three independent biological replicates.

    Journal: Frontiers in Plant Science

    Article Title: BhbZIP60 from Resurrection Plant Boea hygrometrica Is an mRNA Splicing-Activated Endoplasmic Reticulum Stress Regulator Involved in Drought Tolerance

    doi: 10.3389/fpls.2017.00245

    Figure Lengend Snippet: Complementation experiment of Arabidopsis atbzip60 mutant. Complementation of atbzip60 by BhbZIP60S was demonstrated by the expression of UPR genes detected using qRT-PCR. OE(atbzip60)-1 and OE(atbzip60)-2 represent overexpressing BhbZIP60S in atbzip60 background. 18S rRNA was used as an internal reference gene. Data represent the means ± SD of three independent biological replicates.

    Article Snippet: Semi-Quantitative RT-PCR (RT-PCR) and Quantitative Real-Time PCR (qRT-PCR) Analysis Total RNA was extracted using Trizol reagent (TaKaRa).

    Techniques: Mutagenesis, Expressing, Quantitative RT-PCR

    Expression of BhbZIP60S activated ER-QC genes and ABA responsive genes. (A) qRT-PCR analysis of the expression of ER-QC genes. RNA was extracted from seedlings growing on the 0.5 × MS medium with 0, 0.5 μg ml -1 TM or 400 mM mannitol for 3 weeks. (B) qRT-PCR analysis of the expression of drought-responsive genes. RNA was extracted from leaves of 4-week-old plants that were air-dried for 0 h (control), 3 h, and 6 h. Data represent means ± SD of three independent biological replicates.

    Journal: Frontiers in Plant Science

    Article Title: BhbZIP60 from Resurrection Plant Boea hygrometrica Is an mRNA Splicing-Activated Endoplasmic Reticulum Stress Regulator Involved in Drought Tolerance

    doi: 10.3389/fpls.2017.00245

    Figure Lengend Snippet: Expression of BhbZIP60S activated ER-QC genes and ABA responsive genes. (A) qRT-PCR analysis of the expression of ER-QC genes. RNA was extracted from seedlings growing on the 0.5 × MS medium with 0, 0.5 μg ml -1 TM or 400 mM mannitol for 3 weeks. (B) qRT-PCR analysis of the expression of drought-responsive genes. RNA was extracted from leaves of 4-week-old plants that were air-dried for 0 h (control), 3 h, and 6 h. Data represent means ± SD of three independent biological replicates.

    Article Snippet: Semi-Quantitative RT-PCR (RT-PCR) and Quantitative Real-Time PCR (qRT-PCR) Analysis Total RNA was extracted using Trizol reagent (TaKaRa).

    Techniques: Expressing, Quantitative RT-PCR, Mass Spectrometry

    Effects of KDZ injection on the expressions of mitochondrial COX I and COX III mRNA in myocardial tissues of rat models of focal cerebral ischemia. Expressions of mitochondrial COX I and COX III mRNA were determined by RT-PCR. GAPDH was used as control. Data are shown as mean ± SEM ( n = 6/group). * P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Kudiezi injection mitigates myocardial injury induced by acute cerebral ischemia in rats

    doi: 10.1186/s12906-016-1514-1

    Figure Lengend Snippet: Effects of KDZ injection on the expressions of mitochondrial COX I and COX III mRNA in myocardial tissues of rat models of focal cerebral ischemia. Expressions of mitochondrial COX I and COX III mRNA were determined by RT-PCR. GAPDH was used as control. Data are shown as mean ± SEM ( n = 6/group). * P

    Article Snippet: Semi-quantitative RT-PCR analysis of COX I and COX III mRNA expressions was performed using the Prime Script RT-PCR kit (Clontech Laboratories Inc., Mountain View, CA, USA) according to the manufacturer’s instructions and a PCR system (AG-22331; Eppendorf, Hamburg, Germany).

    Techniques: Injection, Reverse Transcription Polymerase Chain Reaction

    Overexpression of PP1 increases alternative splicing and promotes tumor cell apoptosis induced by HHT. A stable MCF7 cell line overexpressing PP1 was constructed with a lentiviral expression system. MCF7-Bobi and MCF7-PP1 cells were treated with 0.5 μM HHT. Total RNA was extracted and analyzed by semi-quantitative RT-PCR for the splicing of Bcl-x and Caspase 9. a Overexpression of PP1 in MCF7 cells. b Semi-quantitative RT-PCR analysis of Bcl-x and Caspase 9 splicing from MCF7-Bobi and MCF7-PP1 cells treated with 0.5 μM HHT. c and d Ratio of Bcl-xL/xS and Caspase 9a/9b in MCF7-Bobi and MCF7-PP1 cells (* P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Homoharringtonine regulates the alternative splicing of Bcl-x and caspase 9 through a protein phosphatase 1-dependent mechanism

    doi: 10.1186/s12906-018-2233-6

    Figure Lengend Snippet: Overexpression of PP1 increases alternative splicing and promotes tumor cell apoptosis induced by HHT. A stable MCF7 cell line overexpressing PP1 was constructed with a lentiviral expression system. MCF7-Bobi and MCF7-PP1 cells were treated with 0.5 μM HHT. Total RNA was extracted and analyzed by semi-quantitative RT-PCR for the splicing of Bcl-x and Caspase 9. a Overexpression of PP1 in MCF7 cells. b Semi-quantitative RT-PCR analysis of Bcl-x and Caspase 9 splicing from MCF7-Bobi and MCF7-PP1 cells treated with 0.5 μM HHT. c and d Ratio of Bcl-xL/xS and Caspase 9a/9b in MCF7-Bobi and MCF7-PP1 cells (* P

    Article Snippet: Semi-quantitative RT-PCR Total RNA was extracted from cultured MCF7, A549 and UACC903 cells using Trizol reagent (Takara, Shiga, Japan) according to the manufacturer’s instruction.

    Techniques: Over Expression, Construct, Expressing, Quantitative RT-PCR

    HHT regulates Bcl-x and Caspase 9 splicing in MCF7 cells. MCF7 cells were treated with HHT. Total RNA was extracted and analyzed by semi-quantitative RT-PCR for the alternative splicing of Bcl-x and Caspase 9. a Decrease of Bcl-xL and increase of Bcl-xS is correlated with HHT concentration. b Densitometric analysis of the ratio of Bcl-xL/xS (* P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Homoharringtonine regulates the alternative splicing of Bcl-x and caspase 9 through a protein phosphatase 1-dependent mechanism

    doi: 10.1186/s12906-018-2233-6

    Figure Lengend Snippet: HHT regulates Bcl-x and Caspase 9 splicing in MCF7 cells. MCF7 cells were treated with HHT. Total RNA was extracted and analyzed by semi-quantitative RT-PCR for the alternative splicing of Bcl-x and Caspase 9. a Decrease of Bcl-xL and increase of Bcl-xS is correlated with HHT concentration. b Densitometric analysis of the ratio of Bcl-xL/xS (* P

    Article Snippet: Semi-quantitative RT-PCR Total RNA was extracted from cultured MCF7, A549 and UACC903 cells using Trizol reagent (Takara, Shiga, Japan) according to the manufacturer’s instruction.

    Techniques: Quantitative RT-PCR, Concentration Assay

    HHT regulates Bcl-x and Caspase 9 splicing in A549 and UACC903 cells. Total RNA was extracted from A549 cells and UACC903 cells treated with HHT. The alternative splicing of Bcl-x and Caspase 9 was then analyzed by semi-quantitative RT-PCR. a Semi-quantitative RT-PCR analysis of Bcl-x splicing from A549 cells treated with different concentrations of HHT. b Densitometric analysis of the ratio of Bcl-xL/xS in A549 cells treated by HHT (* P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Homoharringtonine regulates the alternative splicing of Bcl-x and caspase 9 through a protein phosphatase 1-dependent mechanism

    doi: 10.1186/s12906-018-2233-6

    Figure Lengend Snippet: HHT regulates Bcl-x and Caspase 9 splicing in A549 and UACC903 cells. Total RNA was extracted from A549 cells and UACC903 cells treated with HHT. The alternative splicing of Bcl-x and Caspase 9 was then analyzed by semi-quantitative RT-PCR. a Semi-quantitative RT-PCR analysis of Bcl-x splicing from A549 cells treated with different concentrations of HHT. b Densitometric analysis of the ratio of Bcl-xL/xS in A549 cells treated by HHT (* P

    Article Snippet: Semi-quantitative RT-PCR Total RNA was extracted from cultured MCF7, A549 and UACC903 cells using Trizol reagent (Takara, Shiga, Japan) according to the manufacturer’s instruction.

    Techniques: Quantitative RT-PCR

    Transcriptional analysis of genes upstream and downstream from five clonal mutants. Genomic DNA-free total RNA from wild-type E . chaffeensis (lane 1) and clonal mutants (lane 2) was assessed for the genes 5’ and 3’ to insertion sites by semi quantitative RT-PCR. Panels 1A-5A have the cartoons depicting the insertions and their orientations within the genome and the genes near the insertions. The annotated/predicted gene names near the insertions were provided in each sub-panel. Panels 1B-5B had RT-PCR products resolved on agarose gels for each of the respective genes as in panels 1A-5A. No template controls (lane 3) and wild type genomic DNA template reactions (lane 4) were included to serve as negative and positive controls for each reaction, respectively. M represents 1 kb plus DNA ladder (Life Technologies. Carlsbad, CA). Molecular sizes of the resolved DNA ladder shown in the Figs are the same for each sub-panels and were identified in the left image of panel 1B. Predicted amplicon for each gene was listed in Table 1 . The primer binding locations relative to whole genome data were also provided in Table 1 .

    Journal: PLoS ONE

    Article Title: Mutations in Ehrlichia chaffeensis Causing Polar Effects in Gene Expression and Differential Host Specificities

    doi: 10.1371/journal.pone.0132657

    Figure Lengend Snippet: Transcriptional analysis of genes upstream and downstream from five clonal mutants. Genomic DNA-free total RNA from wild-type E . chaffeensis (lane 1) and clonal mutants (lane 2) was assessed for the genes 5’ and 3’ to insertion sites by semi quantitative RT-PCR. Panels 1A-5A have the cartoons depicting the insertions and their orientations within the genome and the genes near the insertions. The annotated/predicted gene names near the insertions were provided in each sub-panel. Panels 1B-5B had RT-PCR products resolved on agarose gels for each of the respective genes as in panels 1A-5A. No template controls (lane 3) and wild type genomic DNA template reactions (lane 4) were included to serve as negative and positive controls for each reaction, respectively. M represents 1 kb plus DNA ladder (Life Technologies. Carlsbad, CA). Molecular sizes of the resolved DNA ladder shown in the Figs are the same for each sub-panels and were identified in the left image of panel 1B. Predicted amplicon for each gene was listed in Table 1 . The primer binding locations relative to whole genome data were also provided in Table 1 .

    Article Snippet: Semi-quantitative one-step RT-PCR (Life Technologies, Carlsbad, CA) targeting to E . chaffeensis genes surrounding the transposon insertion sites was performed with 35 cycles of amplification [ ] using the gene specific primer sets ( ).

    Techniques: Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Amplification, Binding Assay

    Theoretical models of CFTR mRNA fragments and RNA folding assay results. A , WT CFTR fragment model. B , ΔF508 (Ile507ATT) CFTR fragment model. C , variant (Ile507ATC) ΔF508 CFTR fragment model. Ile-507 ( red ); the CTT deletion site (CUU) is indicated by brackets ; the structures that are identical to the full-length mRNA models are highlighted in gray . Nucleotides corresponding to the full-length sequences are labeled green ; blue lines represent the projected PCR products predicted following T1 RNase digestion; single-stranded Gs (T1 site) are indicated by X ; primers 1 and 2: complementary RT-PCR primer sequences used to amplify WT or ΔF508 (Ile507ATT) CFTR-specific products. D , RT-PCR amplification of WT and ΔF508 (Ile507ATT) CFTR-specific mRNA fragments following RNase T1 digestion. Arrows indicate WT (61 bp) ΔF508 (55 bp) fragment structure specific PCR products in samples digested with 0.05 units of RNase T1. M : molecular weight marker. A representative gel of four individual experiments is shown.

    Journal: The Journal of Biological Chemistry

    Article Title: A Synonymous Single Nucleotide Polymorphism in ?F508 CFTR Alters the Secondary Structure of the mRNA and the Expression of the Mutant Protein *

    doi: 10.1074/jbc.M110.154575

    Figure Lengend Snippet: Theoretical models of CFTR mRNA fragments and RNA folding assay results. A , WT CFTR fragment model. B , ΔF508 (Ile507ATT) CFTR fragment model. C , variant (Ile507ATC) ΔF508 CFTR fragment model. Ile-507 ( red ); the CTT deletion site (CUU) is indicated by brackets ; the structures that are identical to the full-length mRNA models are highlighted in gray . Nucleotides corresponding to the full-length sequences are labeled green ; blue lines represent the projected PCR products predicted following T1 RNase digestion; single-stranded Gs (T1 site) are indicated by X ; primers 1 and 2: complementary RT-PCR primer sequences used to amplify WT or ΔF508 (Ile507ATT) CFTR-specific products. D , RT-PCR amplification of WT and ΔF508 (Ile507ATT) CFTR-specific mRNA fragments following RNase T1 digestion. Arrows indicate WT (61 bp) ΔF508 (55 bp) fragment structure specific PCR products in samples digested with 0.05 units of RNase T1. M : molecular weight marker. A representative gel of four individual experiments is shown.

    Article Snippet: Following a 70% ethanol wash and air-drying, samples containing the RNA digests were solubilized in H2 O and used in one-step semi-quantitative RT-PCR (Applied Biosystems 4309169) to amplify WT and ΔF508 CFTR-specific fragments.

    Techniques: Variant Assay, Labeling, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Amplification, Molecular Weight, Marker

    CFTR mRNA half-life and translational rate measurements. A , WT and ΔF508 CFTR mRNA half-life measurements. Cells expressing HeLaWT and HeLaΔF cells were treated with actinomycin D to inhibit transcription. RNA was isolated at the time points specified and relative CFTR mRNA levels were measured by real time RT-PCR. CFTR mRNA levels at the start of the experiment ( T 0 ) were set as 100%. Results are plotted as % of CFTR mRNA/t 0 . CFTR mRNA half-lives were calculated from the exponential decay based on trend line equation C/C0 = e − k d t (C and C 0 are mRNA amounts after time t and t 0 , respectively, and k d is the mRNA decay constant; R 2 = 0.99). Results were validated in three independent experiments. B , in vitro CFTR translation rates in the presence of microsomes and ATA. ATA was added 10 min after initiation of translation. Incorporation of [ 35 S]methionine was measured 15, 20, 25, 30, 50, and 60 min following addition of ATA. *, significant differences were found in 35 S incorporation levels between WT and ΔF508 (Ile507ATT) CFTR translation at 15, 20, 25, and 30 min; n = 6; p

    Journal: The Journal of Biological Chemistry

    Article Title: A Synonymous Single Nucleotide Polymorphism in ?F508 CFTR Alters the Secondary Structure of the mRNA and the Expression of the Mutant Protein *

    doi: 10.1074/jbc.M110.154575

    Figure Lengend Snippet: CFTR mRNA half-life and translational rate measurements. A , WT and ΔF508 CFTR mRNA half-life measurements. Cells expressing HeLaWT and HeLaΔF cells were treated with actinomycin D to inhibit transcription. RNA was isolated at the time points specified and relative CFTR mRNA levels were measured by real time RT-PCR. CFTR mRNA levels at the start of the experiment ( T 0 ) were set as 100%. Results are plotted as % of CFTR mRNA/t 0 . CFTR mRNA half-lives were calculated from the exponential decay based on trend line equation C/C0 = e − k d t (C and C 0 are mRNA amounts after time t and t 0 , respectively, and k d is the mRNA decay constant; R 2 = 0.99). Results were validated in three independent experiments. B , in vitro CFTR translation rates in the presence of microsomes and ATA. ATA was added 10 min after initiation of translation. Incorporation of [ 35 S]methionine was measured 15, 20, 25, 30, 50, and 60 min following addition of ATA. *, significant differences were found in 35 S incorporation levels between WT and ΔF508 (Ile507ATT) CFTR translation at 15, 20, 25, and 30 min; n = 6; p

    Article Snippet: Following a 70% ethanol wash and air-drying, samples containing the RNA digests were solubilized in H2 O and used in one-step semi-quantitative RT-PCR (Applied Biosystems 4309169) to amplify WT and ΔF508 CFTR-specific fragments.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, In Vitro

    PEA attenuates Aβ-induced astrocyte activation. Aβ-challenged (1 μg/ml) cells were treated with PEA (10 −7 M) in the presence of the selective PPAR-γ antagonist (GW9662, 9 nM) or the selective PPAR-α antagonist (MK886, 3 μM). GFAP mRNA was evaluated 12 hrs following Aβ challenge; GFAP and S100B protein expression was evaluated after 24 hrs of treatments by Western blot and immunofluorescence analyses. S100B release in the cellular milieu was determined 24 hrs after Aβ challenge by ELISA assay. (A) Results of GFAP and S100B Western blot analysis and densitometric analysis of corresponding bands. β-actin was used as loading control. (B) Results of GFAP RT-PCR amplification and densitometric analysis of corresponding bands. GAPDH was used as standard control. (C) Measurement of S100B release by ELISA assay. (D) Analysis of GFAP and S100B protein expression by immunofluorescence (magnification 10 × ). Results are the mean ± S.E.M. of n = 4 separate experiments. *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Palmitoylethanolamide counteracts reactive astrogliosis induced by β-amyloid peptide

    doi: 10.1111/j.1582-4934.2011.01267.x

    Figure Lengend Snippet: PEA attenuates Aβ-induced astrocyte activation. Aβ-challenged (1 μg/ml) cells were treated with PEA (10 −7 M) in the presence of the selective PPAR-γ antagonist (GW9662, 9 nM) or the selective PPAR-α antagonist (MK886, 3 μM). GFAP mRNA was evaluated 12 hrs following Aβ challenge; GFAP and S100B protein expression was evaluated after 24 hrs of treatments by Western blot and immunofluorescence analyses. S100B release in the cellular milieu was determined 24 hrs after Aβ challenge by ELISA assay. (A) Results of GFAP and S100B Western blot analysis and densitometric analysis of corresponding bands. β-actin was used as loading control. (B) Results of GFAP RT-PCR amplification and densitometric analysis of corresponding bands. GAPDH was used as standard control. (C) Measurement of S100B release by ELISA assay. (D) Analysis of GFAP and S100B protein expression by immunofluorescence (magnification 10 × ). Results are the mean ± S.E.M. of n = 4 separate experiments. *** P

    Article Snippet: Reverse transcriptase-PCR analysis The mRNA level was determined using the semi-quantitative RT-PCR method (Invitrogen).

    Techniques: Activation Assay, Expressing, Western Blot, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Amplification

    Fig. 5. Quantitation of the hypertrophic phenotype of AdMEK1-infected cardiomyocytes. ( A ) Cell surface area was measured in α-actinin-stained cardiomyocyte cultures using confocal microscopy and digitized imaging (24 h after infection). Cultures were left in serum-free medium with no stimulation or were infected with Adβgal or AdMEK1, or were treated with U0126. ( B ) Cardiomyocytes were also stained with an ANF-specific antibody to quantify the percentage of cells expressing ANF. The data in (A) and (B) were obtained in two independent experiments. ( C ) Total ANF mRNA levels were also quantified by RT–PCR, showing similar induction of ANF mRNA between 1% FBS and AdMEK1 infection. ( D ) Protein synthesis rates were monitored by the incorporation of [ 3 H]leucine in Adβgal-, AdMEK1- and 1% FBS-stimulated cardiomyocytes. * P

    Journal: The EMBO Journal

    Article Title: The MEK1-ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice

    doi: 10.1093/emboj/19.23.6341

    Figure Lengend Snippet: Fig. 5. Quantitation of the hypertrophic phenotype of AdMEK1-infected cardiomyocytes. ( A ) Cell surface area was measured in α-actinin-stained cardiomyocyte cultures using confocal microscopy and digitized imaging (24 h after infection). Cultures were left in serum-free medium with no stimulation or were infected with Adβgal or AdMEK1, or were treated with U0126. ( B ) Cardiomyocytes were also stained with an ANF-specific antibody to quantify the percentage of cells expressing ANF. The data in (A) and (B) were obtained in two independent experiments. ( C ) Total ANF mRNA levels were also quantified by RT–PCR, showing similar induction of ANF mRNA between 1% FBS and AdMEK1 infection. ( D ) Protein synthesis rates were monitored by the incorporation of [ 3 H]leucine in Adβgal-, AdMEK1- and 1% FBS-stimulated cardiomyocytes. * P

    Article Snippet: Analysis of mRNA levels for ANF and L7 (ribosomal protein control) from cardiomyocyte cultures was performed by semi-quantitative RT–PCR (Titan one tube RT–PCR; Boehringer Mannheim) in the presence of [γ-32 P]dCTP with primers designed to detect rat gene products exactly as described previously ( ).

    Techniques: Quantitation Assay, Infection, Staining, Confocal Microscopy, Imaging, Expressing, Reverse Transcription Polymerase Chain Reaction

    RT–PCR of AQP4 of striatum (a), and of cortex (b) of different set of experimental rats. M denotes marker i.e. DNA ladder. C denotes negative control. S denotes sham groups, S+D denote sham treated with drug, V denotes the vehicle and V+D denote the vehicle treated with drug. Histograms represent cumulative data expressed as mean ± SEM obtained from three different sets of experiments conducted for striatum and cortex. ( a vs sham b vs vehicle) ( P ≤0.05).

    Journal: PLoS ONE

    Article Title: Aquaporin-4 Inhibition Mediates Piroxicam-Induced Neuroprotection against Focal Cerebral Ischemia/Reperfusion Injury in Rodents

    doi: 10.1371/journal.pone.0073481

    Figure Lengend Snippet: RT–PCR of AQP4 of striatum (a), and of cortex (b) of different set of experimental rats. M denotes marker i.e. DNA ladder. C denotes negative control. S denotes sham groups, S+D denote sham treated with drug, V denotes the vehicle and V+D denote the vehicle treated with drug. Histograms represent cumulative data expressed as mean ± SEM obtained from three different sets of experiments conducted for striatum and cortex. ( a vs sham b vs vehicle) ( P ≤0.05).

    Article Snippet: Semi- quantitative RT-PCR of AQP4 For getting amplification in linear range for semi-quantitative RT–PCR, the number of cycles considered for AQP4 was 29 and for β-actin, it was considered 26 as previously reported by Gupta et al., (2011) .

    Techniques: Reverse Transcription Polymerase Chain Reaction, Marker, Negative Control

    (A) qRT-PCR amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no miRNA was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Conservation of a microRNA cluster in parasitic nematodes and profiling of miRNAs in excretory-secretory products and microvesicles of Haemonchus contortus

    doi: 10.1371/journal.pntd.0006056

    Figure Lengend Snippet: (A) qRT-PCR amplification of Hco-miR-5352 in adult female whole worms, adult ES and adult gut tissue. Results shown represent the mean ± SD of three technical replicates. Hco-miR-50 was used as a control gene for PCR amplification. Raw qRT-PCR readings are shown as no miRNA was known that could be used to normalise expression across all samples. (B) qRT-PCR amplification analysis of Hco-miR-50 and Hco-miR-5352 expression in adult females and egg/L1 sample. Results shown represent the mean ± SD of three technical replicates, with Hco-mir-50 shown as a control for PCR amplification as used in Fig 3A.

    Article Snippet: Semi-quantitative RT-PCR RT-PCR was performed using the miRNA qRT-PCR Master Mix protocol (Agilent Technologies) on RNA extracted from whole adult worms, adult worm ES or abomasal and lymph node tissue from infected or uninfected sheep.

    Techniques: Quantitative RT-PCR, Amplification, Polymerase Chain Reaction, Expressing

    qRT-PCR to detect presence of A) Hco-miR-5352 , B) Hco-miR-5895 and C) Hco-miR-5960 in two infected and two pathogen-free (uninfected) sheep from abomasal (abo) and draining lymph node (lymph) tissue. Samples were obtained from the abomasum and lymph nodes and qRT-PCR was carried out in triplicate. Mean values are shown with error bars representing the SD. Fluorescence values were normalised to oar-miR-26a qRT-PCR. P values shown were calculated using an analysis of variance test.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Conservation of a microRNA cluster in parasitic nematodes and profiling of miRNAs in excretory-secretory products and microvesicles of Haemonchus contortus

    doi: 10.1371/journal.pntd.0006056

    Figure Lengend Snippet: qRT-PCR to detect presence of A) Hco-miR-5352 , B) Hco-miR-5895 and C) Hco-miR-5960 in two infected and two pathogen-free (uninfected) sheep from abomasal (abo) and draining lymph node (lymph) tissue. Samples were obtained from the abomasum and lymph nodes and qRT-PCR was carried out in triplicate. Mean values are shown with error bars representing the SD. Fluorescence values were normalised to oar-miR-26a qRT-PCR. P values shown were calculated using an analysis of variance test.

    Article Snippet: Semi-quantitative RT-PCR RT-PCR was performed using the miRNA qRT-PCR Master Mix protocol (Agilent Technologies) on RNA extracted from whole adult worms, adult worm ES or abomasal and lymph node tissue from infected or uninfected sheep.

    Techniques: Quantitative RT-PCR, Infection, Fluorescence

    JIKI trial mortality, according to age and baseline RT-PCR Ct value. Histograms represent mortality percentages. Vertical bars represent 95% confidence intervals. Red bars represent target values. The RT-PCR assay was conducted using the RealStar Filovirus Screen RT-PCR Kit 1.0 (Altona Diagnostics).

    Journal: PLoS Medicine

    Article Title: Experimental Treatment with Favipiravir for Ebola Virus Disease (the JIKI Trial): A Historically Controlled, Single-Arm Proof-of-Concept Trial in Guinea

    doi: 10.1371/journal.pmed.1001967

    Figure Lengend Snippet: JIKI trial mortality, according to age and baseline RT-PCR Ct value. Histograms represent mortality percentages. Vertical bars represent 95% confidence intervals. Red bars represent target values. The RT-PCR assay was conducted using the RealStar Filovirus Screen RT-PCR Kit 1.0 (Altona Diagnostics).

    Article Snippet: Initial diagnosis of EVD was made using a semi-quantitative RT-PCR assay (RealStar Filovirus Screen RT-PCR Kit 1.0, Altona Diagnostics).

    Techniques: Reverse Transcription Polymerase Chain Reaction

    Tissue-restricted expression of ELA in the human. Reverse transcription PCR was conducted on total RNAs extracted from the indicated human tissues, and embryonic (ESCs) and induced (iPSCs) pluripotent stem cells using primers specific for human ELA, APJ and β-actin.

    Journal: Scientific Reports

    Article Title: Elabela-Apelin Receptor Signaling Pathway is Functional in Mammalian Systems

    doi: 10.1038/srep08170

    Figure Lengend Snippet: Tissue-restricted expression of ELA in the human. Reverse transcription PCR was conducted on total RNAs extracted from the indicated human tissues, and embryonic (ESCs) and induced (iPSCs) pluripotent stem cells using primers specific for human ELA, APJ and β-actin.

    Article Snippet: Semi-quantitative RT-PCR Tissue total RNAs were purchased from Clontech.

    Techniques: Expressing, Polymerase Chain Reaction

    qRT-PCR analysis of Heat shock protein70 gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P

    Journal: PLoS ONE

    Article Title: Expression of Heat Shock Protein 70 Gene and Its Correlation with Inflammatory Markers in Essential Hypertension

    doi: 10.1371/journal.pone.0151060

    Figure Lengend Snippet: qRT-PCR analysis of Heat shock protein70 gene expressions at mRNA level and indirect ELISA analysis at protein level. (A) represents the scattered plot of levels of HSP70 gene expression at mRNA level in the individual study subjects by qRT-PCR. The relative HSP70 gene expression in patients and controls is expressed in delta-CT values. Results were expressed as densitometric ratio (HSP70 to 18S rRNA gene) in patients and controls group ± SD. P

    Article Snippet: Semi-quantitative PCR Semi-quantitative PCR for heat shock protein70 (HSP70) gene was performed by using Mastercycler (eppendorf).

    Techniques: Quantitative RT-PCR, Indirect ELISA, Expressing

    In vivo NHP infection with B. burgdorferi increases NK-1R expression in the CNS, and such increases are prevented by treatment with the NK-1R antagonist aprepitant. Rhesus macaques were uninfected ( n = 2 animals) or infected intrathecally with B. burgdorferi (Bb, 1 × 10 8 bacteria; n = 8), and infected animals were either untreated ( n = 4) or treated with aprepitant ( n = 4) for 2 or 4 weeks prior to euthanasia. Expression of mRNA encoding NK-1R in frontal cortical tissue samples was determined by RT-PCR ( a ) and relative expression normalized to GAPDH levels was determined by densitometric analysis ( b ). NK-1R protein expression was determined in tissue samples by immunoblot analysis and normalized to β-actin expression ( c ). Data is expressed as the mean ± SD. Asterisk and pound symbols indicate statistically significant difference from uninfected animals and untreated infected animals, respectively ( p

    Journal: Journal of Neuroinflammation

    Article Title: Aprepitant limits in vivo neuroinflammatory responses in a rhesus model of Lyme neuroborreliosis

    doi: 10.1186/s12974-017-0813-x

    Figure Lengend Snippet: In vivo NHP infection with B. burgdorferi increases NK-1R expression in the CNS, and such increases are prevented by treatment with the NK-1R antagonist aprepitant. Rhesus macaques were uninfected ( n = 2 animals) or infected intrathecally with B. burgdorferi (Bb, 1 × 10 8 bacteria; n = 8), and infected animals were either untreated ( n = 4) or treated with aprepitant ( n = 4) for 2 or 4 weeks prior to euthanasia. Expression of mRNA encoding NK-1R in frontal cortical tissue samples was determined by RT-PCR ( a ) and relative expression normalized to GAPDH levels was determined by densitometric analysis ( b ). NK-1R protein expression was determined in tissue samples by immunoblot analysis and normalized to β-actin expression ( c ). Data is expressed as the mean ± SD. Asterisk and pound symbols indicate statistically significant difference from uninfected animals and untreated infected animals, respectively ( p

    Article Snippet: Semi-quantitative reverse transcription-PCR and quantitative real-time PCR Promega flexi was utilized in semi quantitative reverse transcription-PCR (RT-PCR) to assess levels of mRNA encoding NK-1R as previously described [ , ], and quantification was performed using Bio-Rad ImageLab software and normalized to the expression of GAPDH determined in parallel RT-PCR reactions.

    Techniques: In Vivo, Infection, Expressing, Reverse Transcription Polymerase Chain Reaction

    Clustering of the differentially expressed proteins and a comparison of the expression patterns for ten proteins and their transcripts. The hierarchical clustering of the abundance volume of the protein spots ( A ) and their relative volume ratios ( B ) at S3 on 2-DE gels are shown. The average changed abundance ratios of the identified protein spots on 2-D DIGE gels are also presented ( C ). The high- or low-abundance protein spots or the induced- or reduced-values for the relative changed ratios were indicated in red or green, respectively. The details for hierarchical clustering of the changed patterns of the differential protein spots on 2-DE and 2-D DIGE are presented in Table S2 and Table S3 , respectively. Then, 10 DEPs with sharp changes were selected to determine the correlations between their gene and protein expression levels using RT-PCR and qRT-PCR. The spot numbers for a specific protein are given on the right side of the heat plot next to the same protein in the heat plot. For each treatment, the expression of each gene is presented as a ratio between the gene value and the corresponding 18SRNA value. The gray dot line in each rectangle graph represents a 1.0 ratio value. The error bars indicate the SD of three independent PCR reactions for three biological replicates. For UDXS and MADH, antibodies were used to determine the protein expression, revealing that the two proteins had similar expression patterns to those on the 2-DE gels ( D ). Abbreviations: AGPS, ADP-glucose pyrophosphorylase small subunit; GLUP, glucan phosphorylase; APX1, ascorbate peroxidase-1; initiation factor eIF5-A; SOD, superoxide dismutase; SAMS, S-adenosylmethionine synthetase; UDXS, UDP-D-apiose/UDP-D-xylose synthase; MADH, malate dehydrogenase; WB, Western blotting.

    Journal: Scientific Reports

    Article Title: Proteomics Profiling Reveals Carbohydrate Metabolic Enzymes and 14-3-3 Proteins Play Important Roles for Starch Accumulation during Cassava Root Tuberization

    doi: 10.1038/srep19643

    Figure Lengend Snippet: Clustering of the differentially expressed proteins and a comparison of the expression patterns for ten proteins and their transcripts. The hierarchical clustering of the abundance volume of the protein spots ( A ) and their relative volume ratios ( B ) at S3 on 2-DE gels are shown. The average changed abundance ratios of the identified protein spots on 2-D DIGE gels are also presented ( C ). The high- or low-abundance protein spots or the induced- or reduced-values for the relative changed ratios were indicated in red or green, respectively. The details for hierarchical clustering of the changed patterns of the differential protein spots on 2-DE and 2-D DIGE are presented in Table S2 and Table S3 , respectively. Then, 10 DEPs with sharp changes were selected to determine the correlations between their gene and protein expression levels using RT-PCR and qRT-PCR. The spot numbers for a specific protein are given on the right side of the heat plot next to the same protein in the heat plot. For each treatment, the expression of each gene is presented as a ratio between the gene value and the corresponding 18SRNA value. The gray dot line in each rectangle graph represents a 1.0 ratio value. The error bars indicate the SD of three independent PCR reactions for three biological replicates. For UDXS and MADH, antibodies were used to determine the protein expression, revealing that the two proteins had similar expression patterns to those on the 2-DE gels ( D ). Abbreviations: AGPS, ADP-glucose pyrophosphorylase small subunit; GLUP, glucan phosphorylase; APX1, ascorbate peroxidase-1; initiation factor eIF5-A; SOD, superoxide dismutase; SAMS, S-adenosylmethionine synthetase; UDXS, UDP-D-apiose/UDP-D-xylose synthase; MADH, malate dehydrogenase; WB, Western blotting.

    Article Snippet: Semi-quantitative reverse transcription PCR (RT-PCR) and quantitative RT-PCR (qRT-PCR) analysis Total RNA was used to generate cDNAs using the reverse transcriptase kit reagents (TaKaRa, Tokyo, Japan).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Polymerase Chain Reaction, Western Blot

    Determination of the induced changes of the 14-3-3 protein in both cassava tuberous roots and transgenic Arabidopsis plants. The typical area of the 14-3-3 isoforms on the 2-D DIGE gel ( A ) and the merged Pro-Q Diamond- and SYPRO Ruby-stained gels ( B ) are highlighted. Abundance clusters of the three identified 14-3-3 protein spots ( C ) at different tuberous root developmental stages (S2-S7) are also presented. The total protein abundance detected by Western blotting ( D ) and the general gene expression detected by RT-PCR ( E ) and qRT-PCR ( F ) demonstrated similar changes in the expression patterns during tuberous root development. Three lines (L1-3) of hygromycin-resistant ( G ) transgenic Arabidopsis plants were selected to produce T3 generation seeds. In standard MS medium ( H ), the T3 seedlings showed no difference compared to the wild type (WT) or the line overexpressing the P-super 1300 + vector (LV). PCR analysis verified that the cassava 14-3-3 gene was subcloned into the genomes of the three lines, but not into the WT or LV plants ( I ). RT-PCR ( J ) and qRT-PCR ( L ) revealed that the target gene had different expression levels. The Arabidopsis actin gene was used as a reference ( K ). The fragment number of T-DNA insertions was determined by Southern blotting ( M ). The protein abundances in the different plant lines were determined by Western blotting using a polyclonal antibody against cassava 14-3-3 protein ( N ). Starch accumulation in the leaves of different plants was visualized using an iodine solution ( O ). The soluble sugar ( P,Q ) and starch ( R,S ) concentrations in both the leaves ( P,R ) and roots ( Q,S ) of the different Arabidopsis lines (WT, LV, and L1-3) were also determined. The gray dot line in each qRT-PCR rectangle graph represents a 1.0 ratio value. In figures P–S , the columns labeled with a and b or b and c indicate significant difference (p

    Journal: Scientific Reports

    Article Title: Proteomics Profiling Reveals Carbohydrate Metabolic Enzymes and 14-3-3 Proteins Play Important Roles for Starch Accumulation during Cassava Root Tuberization

    doi: 10.1038/srep19643

    Figure Lengend Snippet: Determination of the induced changes of the 14-3-3 protein in both cassava tuberous roots and transgenic Arabidopsis plants. The typical area of the 14-3-3 isoforms on the 2-D DIGE gel ( A ) and the merged Pro-Q Diamond- and SYPRO Ruby-stained gels ( B ) are highlighted. Abundance clusters of the three identified 14-3-3 protein spots ( C ) at different tuberous root developmental stages (S2-S7) are also presented. The total protein abundance detected by Western blotting ( D ) and the general gene expression detected by RT-PCR ( E ) and qRT-PCR ( F ) demonstrated similar changes in the expression patterns during tuberous root development. Three lines (L1-3) of hygromycin-resistant ( G ) transgenic Arabidopsis plants were selected to produce T3 generation seeds. In standard MS medium ( H ), the T3 seedlings showed no difference compared to the wild type (WT) or the line overexpressing the P-super 1300 + vector (LV). PCR analysis verified that the cassava 14-3-3 gene was subcloned into the genomes of the three lines, but not into the WT or LV plants ( I ). RT-PCR ( J ) and qRT-PCR ( L ) revealed that the target gene had different expression levels. The Arabidopsis actin gene was used as a reference ( K ). The fragment number of T-DNA insertions was determined by Southern blotting ( M ). The protein abundances in the different plant lines were determined by Western blotting using a polyclonal antibody against cassava 14-3-3 protein ( N ). Starch accumulation in the leaves of different plants was visualized using an iodine solution ( O ). The soluble sugar ( P,Q ) and starch ( R,S ) concentrations in both the leaves ( P,R ) and roots ( Q,S ) of the different Arabidopsis lines (WT, LV, and L1-3) were also determined. The gray dot line in each qRT-PCR rectangle graph represents a 1.0 ratio value. In figures P–S , the columns labeled with a and b or b and c indicate significant difference (p

    Article Snippet: Semi-quantitative reverse transcription PCR (RT-PCR) and quantitative RT-PCR (qRT-PCR) analysis Total RNA was used to generate cDNAs using the reverse transcriptase kit reagents (TaKaRa, Tokyo, Japan).

    Techniques: Transgenic Assay, Staining, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Mass Spectrometry, Plasmid Preparation, Polymerase Chain Reaction, Southern Blot, Labeling