qrt pcr analysis Takara Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Thermo Fisher 7500 fast real time pcr system
    7500 Fast Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 32786 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7500 fast real time pcr system/product/Thermo Fisher
    Average 90 stars, based on 32786 article reviews
    Price from $9.99 to $1999.99
    7500 fast real time pcr system - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    77
    TaKaRa polymerase chain reaction qrt pcr analysis
    Zebrafish bach1b is a co-ortholog of mammalian Bach1 , and heme regulates exocrine peptidase precursor genes. (A) Zebrafish Bach1b contains four CP motifs, three of which are also found in human and mouse. (B) Phylogenetic analysis of Bach1 proteins shows that zebrafish contain two bach1 genes, bach1a and bach1b , which are co-orthologs of mammalian Bach1 . The tree was constructed using the neighbor-joining method with MEGA5 ( Tamura et al., 2011 ). The numbers indicate the percentage bootstrap support. Dr , Danio rerio; Tr , Takifugu rubripes; Tn , Tetraodon nigroviridis; Ol , Oryzias latipes; Ga , Gasterosteus aculeatus; On , Oreochromis niloticus; Xt , Xenopus tropicalis; Ci , Ciona intestinalis; Cs , Ciona savignyi; Rn , Rattus norvegicus; Mm , Mus musculus; Hs , Homo sapiens. Ciona intestinalis and Ciona savignyi Bach proteins were used as an outgroup. The Ensembl gene IDs of these genes are listed in supplementary material Table S2 . (C) Downregulation of six peptidase precursor genes in zebrafish yquem/urod (−/−) and their rescue upon treatment with hemin. The concentration of hemin used was 100 μM. The expression levels of six peptidase precursor genes in larvae at 84 hpf were determined by using <t>qRT-PCR.</t> Student’s t -tests were conducted. * P
    Polymerase Chain Reaction Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr analysis/product/TaKaRa
    Average 77 stars, based on 171 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-02
    77/100 stars
      Buy from Supplier

    81
    TaKaRa real time polymerase chain reaction qrt pcr analyses
    Zebrafish bach1b is a co-ortholog of mammalian Bach1 , and heme regulates exocrine peptidase precursor genes. (A) Zebrafish Bach1b contains four CP motifs, three of which are also found in human and mouse. (B) Phylogenetic analysis of Bach1 proteins shows that zebrafish contain two bach1 genes, bach1a and bach1b , which are co-orthologs of mammalian Bach1 . The tree was constructed using the neighbor-joining method with MEGA5 ( Tamura et al., 2011 ). The numbers indicate the percentage bootstrap support. Dr , Danio rerio; Tr , Takifugu rubripes; Tn , Tetraodon nigroviridis; Ol , Oryzias latipes; Ga , Gasterosteus aculeatus; On , Oreochromis niloticus; Xt , Xenopus tropicalis; Ci , Ciona intestinalis; Cs , Ciona savignyi; Rn , Rattus norvegicus; Mm , Mus musculus; Hs , Homo sapiens. Ciona intestinalis and Ciona savignyi Bach proteins were used as an outgroup. The Ensembl gene IDs of these genes are listed in supplementary material Table S2 . (C) Downregulation of six peptidase precursor genes in zebrafish yquem/urod (−/−) and their rescue upon treatment with hemin. The concentration of hemin used was 100 μM. The expression levels of six peptidase precursor genes in larvae at 84 hpf were determined by using <t>qRT-PCR.</t> Student’s t -tests were conducted. * P
    Real Time Polymerase Chain Reaction Qrt Pcr Analyses, supplied by TaKaRa, used in various techniques. Bioz Stars score: 81/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr analyses/product/TaKaRa
    Average 81 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr analyses - by Bioz Stars, 2020-02
    81/100 stars
      Buy from Supplier

    90
    TaKaRa real time polymerase chain reaction qrt pcr analysis
    Relative mRNA levels of 9 MaAQP genes in BX and FJ were determined by <t>qRT-PCR</t> analysis. ( A – C ) expression patterns of MaPIP2-7, MaPIP1-4 and MaSIP1-1 in different tissues of BX and FJ. The mRNA fold difference was relative to that of BX-root samples used as calibrator; ( D – F ) expression patterns of MaPIP1-6, MaPIP2-10 and MaTIP4-1 in different stages of fruit development and ripening in BX and FJ. The mRNA fold difference was relative to that of BX-0DAF samples used as calibrator; ( G – I ) expression patterns of MaPIP2-6, MaTIP2-1 and MaTIP4-2 in response to cold, salt and osmotic stresses in BX and FJ. The mRNA fold difference was relative to that of untreated samples used as calibrator. Log 2 -based values were used to display differential expression results. Data are means ± SD of n = 3 biological replicates.
    Real Time Polymerase Chain Reaction Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr analysis/product/TaKaRa
    Average 90 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    94
    TaKaRa sybr green real time quantitative polymerase chain reaction qrt pcr analysis
    Relative mRNA levels of 9 MaAQP genes in BX and FJ were determined by <t>qRT-PCR</t> analysis. ( A – C ) expression patterns of MaPIP2-7, MaPIP1-4 and MaSIP1-1 in different tissues of BX and FJ. The mRNA fold difference was relative to that of BX-root samples used as calibrator; ( D – F ) expression patterns of MaPIP1-6, MaPIP2-10 and MaTIP4-1 in different stages of fruit development and ripening in BX and FJ. The mRNA fold difference was relative to that of BX-0DAF samples used as calibrator; ( G – I ) expression patterns of MaPIP2-6, MaTIP2-1 and MaTIP4-2 in response to cold, salt and osmotic stresses in BX and FJ. The mRNA fold difference was relative to that of untreated samples used as calibrator. Log 2 -based values were used to display differential expression results. Data are means ± SD of n = 3 biological replicates.
    Sybr Green Real Time Quantitative Polymerase Chain Reaction Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green real time quantitative polymerase chain reaction qrt pcr analysis/product/TaKaRa
    Average 94 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    sybr green real time quantitative polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-02
    94/100 stars
      Buy from Supplier

    99
    TaKaRa quantitative real time polymerase chain reaction qrt pcr analysis
    Expression patterns of OsCTZFP8 transcript in response to abiotic stresses. <t>qRT-PCR</t> was performed with 2-week-old NT plants exposed to cold (4°C), ABA (5 μ M), and NaCl (250 mM) treatments at different time points. The expression of eEF1α was used as an internal control. Data present the means ± SE of two biological replicates.
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr analysis/product/TaKaRa
    Average 99 stars, based on 73 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    TaKaRa quantitative real time polymerase chain reaction qrt pcr analyses
    Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using <t>qRT-PCR.</t> b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Analyses, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr analyses/product/TaKaRa
    Average 99 stars, based on 42 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr analyses - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    86
    TaKaRa qrt pcr analysis takara minibest universal rna extraction kit
    Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using <t>qRT-PCR.</t> b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P
    Qrt Pcr Analysis Takara Minibest Universal Rna Extraction Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis takara minibest universal rna extraction kit/product/TaKaRa
    Average 86 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis takara minibest universal rna extraction kit - by Bioz Stars, 2020-02
    86/100 stars
      Buy from Supplier

    87
    TaKaRa sybr qrt pcr analysis
    Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using <t>qRT-PCR.</t> b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P
    Sybr Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 87/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr qrt pcr analysis/product/TaKaRa
    Average 87 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    sybr qrt pcr analysis - by Bioz Stars, 2020-02
    87/100 stars
      Buy from Supplier

    83
    TaKaRa qrt pcr pcr
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Qrt Pcr Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 83/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr pcr/product/TaKaRa
    Average 83 stars, based on 123 article reviews
    Price from $9.99 to $1999.99
    qrt pcr pcr - by Bioz Stars, 2020-02
    83/100 stars
      Buy from Supplier

    86
    TaKaRa qrt pcr analysis trizol reagent
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Qrt Pcr Analysis Trizol Reagent, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis trizol reagent/product/TaKaRa
    Average 86 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis trizol reagent - by Bioz Stars, 2020-02
    86/100 stars
      Buy from Supplier

    94
    TaKaRa qrt pcr analysis rnaiso plus
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Qrt Pcr Analysis Rnaiso Plus, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis rnaiso plus/product/TaKaRa
    Average 94 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis rnaiso plus - by Bioz Stars, 2020-02
    94/100 stars
      Buy from Supplier

    99
    TaKaRa sybr green qrt pcr kit
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Sybr Green Qrt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green qrt pcr kit/product/TaKaRa
    Average 99 stars, based on 83 article reviews
    Price from $9.99 to $1999.99
    sybr green qrt pcr kit - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    TaKaRa sybr green qrt pcr analysis
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Sybr Green Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green qrt pcr analysis/product/TaKaRa
    Average 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    sybr green qrt pcr analysis - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    77
    TaKaRa qrt pcr analysis quantitative reverse transcriptase qrt pcr
    <t>qRT-PCR</t> validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.
    Qrt Pcr Analysis Quantitative Reverse Transcriptase Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis quantitative reverse transcriptase qrt pcr/product/TaKaRa
    Average 77 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis quantitative reverse transcriptase qrt pcr - by Bioz Stars, 2020-02
    77/100 stars
      Buy from Supplier

    76
    TaKaRa quantitative real time pcr qrt pcr qrt pcr analyses
    Comparative <t>qRT-PCR</t> analysis of mRNA levels of the Kctd14 gene between Znf230 KO and C57BL/6J (WT) mice. Bars represent the means ± S.D., Statistical p-values
    Quantitative Real Time Pcr Qrt Pcr Qrt Pcr Analyses, supplied by TaKaRa, used in various techniques. Bioz Stars score: 76/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr qrt pcr qrt pcr analyses/product/TaKaRa
    Average 76 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr qrt pcr qrt pcr analyses - by Bioz Stars, 2020-02
    76/100 stars
      Buy from Supplier

    88
    TaKaRa quantitative rt pcr qrt pcr analyses
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Quantitative Rt Pcr Qrt Pcr Analyses, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative rt pcr qrt pcr analyses/product/TaKaRa
    Average 88 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    quantitative rt pcr qrt pcr analyses - by Bioz Stars, 2020-02
    88/100 stars
      Buy from Supplier

    79
    TaKaRa quantitative real time pcr qrt pcr analysis a sybr primix ex taq kit
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Quantitative Real Time Pcr Qrt Pcr Analysis A Sybr Primix Ex Taq Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr qrt pcr analysis a sybr primix ex taq kit/product/TaKaRa
    Average 79 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr qrt pcr analysis a sybr primix ex taq kit - by Bioz Stars, 2020-02
    79/100 stars
      Buy from Supplier

    99
    TaKaRa quantitative real time pcr analysis
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Quantitative Real Time Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 434 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr analysis/product/TaKaRa
    Average 99 stars, based on 434 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr analysis - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    TaKaRa sybr green pcr master mix
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Sybr Green Pcr Master Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 6786 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green pcr master mix/product/TaKaRa
    Average 99 stars, based on 6786 article reviews
    Price from $9.99 to $1999.99
    sybr green pcr master mix - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    90
    Thermo Fisher reaction buffer
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Reaction Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 5752 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reaction buffer/product/Thermo Fisher
    Average 90 stars, based on 5752 article reviews
    Price from $9.99 to $1999.99
    reaction buffer - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    99
    TaKaRa sybr premix ex taq
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Sybr Premix Ex Taq, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 34533 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr premix ex taq/product/TaKaRa
    Average 99 stars, based on 34533 article reviews
    Price from $9.99 to $1999.99
    sybr premix ex taq - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    TaKaRa minibest universal rna extraction kit
    ABI3 regulates the expression of TIP3 genes. (A and B) <t>qRT-PCR</t> and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.
    Minibest Universal Rna Extraction Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1604 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/minibest universal rna extraction kit/product/TaKaRa
    Average 99 stars, based on 1604 article reviews
    Price from $9.99 to $1999.99
    minibest universal rna extraction kit - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    Image Search Results


    Zebrafish bach1b is a co-ortholog of mammalian Bach1 , and heme regulates exocrine peptidase precursor genes. (A) Zebrafish Bach1b contains four CP motifs, three of which are also found in human and mouse. (B) Phylogenetic analysis of Bach1 proteins shows that zebrafish contain two bach1 genes, bach1a and bach1b , which are co-orthologs of mammalian Bach1 . The tree was constructed using the neighbor-joining method with MEGA5 ( Tamura et al., 2011 ). The numbers indicate the percentage bootstrap support. Dr , Danio rerio; Tr , Takifugu rubripes; Tn , Tetraodon nigroviridis; Ol , Oryzias latipes; Ga , Gasterosteus aculeatus; On , Oreochromis niloticus; Xt , Xenopus tropicalis; Ci , Ciona intestinalis; Cs , Ciona savignyi; Rn , Rattus norvegicus; Mm , Mus musculus; Hs , Homo sapiens. Ciona intestinalis and Ciona savignyi Bach proteins were used as an outgroup. The Ensembl gene IDs of these genes are listed in supplementary material Table S2 . (C) Downregulation of six peptidase precursor genes in zebrafish yquem/urod (−/−) and their rescue upon treatment with hemin. The concentration of hemin used was 100 μM. The expression levels of six peptidase precursor genes in larvae at 84 hpf were determined by using qRT-PCR. Student’s t -tests were conducted. * P

    Journal: Disease Models & Mechanisms

    Article Title: Heme acts through the Bach1b/Nrf2a-MafK pathway to regulate exocrine peptidase precursor genes in porphyric zebrafish

    doi: 10.1242/dmm.014951

    Figure Lengend Snippet: Zebrafish bach1b is a co-ortholog of mammalian Bach1 , and heme regulates exocrine peptidase precursor genes. (A) Zebrafish Bach1b contains four CP motifs, three of which are also found in human and mouse. (B) Phylogenetic analysis of Bach1 proteins shows that zebrafish contain two bach1 genes, bach1a and bach1b , which are co-orthologs of mammalian Bach1 . The tree was constructed using the neighbor-joining method with MEGA5 ( Tamura et al., 2011 ). The numbers indicate the percentage bootstrap support. Dr , Danio rerio; Tr , Takifugu rubripes; Tn , Tetraodon nigroviridis; Ol , Oryzias latipes; Ga , Gasterosteus aculeatus; On , Oreochromis niloticus; Xt , Xenopus tropicalis; Ci , Ciona intestinalis; Cs , Ciona savignyi; Rn , Rattus norvegicus; Mm , Mus musculus; Hs , Homo sapiens. Ciona intestinalis and Ciona savignyi Bach proteins were used as an outgroup. The Ensembl gene IDs of these genes are listed in supplementary material Table S2 . (C) Downregulation of six peptidase precursor genes in zebrafish yquem/urod (−/−) and their rescue upon treatment with hemin. The concentration of hemin used was 100 μM. The expression levels of six peptidase precursor genes in larvae at 84 hpf were determined by using qRT-PCR. Student’s t -tests were conducted. * P

    Article Snippet: RNA extraction and qRT-PCR Total RNA was extracted using the TRIzol® Reagent, according to the manufacturer’s instructions (Invitrogen). cDNA was synthesized by using reverse transcription with the M-MLV reverse transcription kit (Invitrogen), which was then used as the template for qRT-PCR analysis. qRT-PCR reactions were performed with the ABI StepOnePlus™ system, using SYBR® Premix Ex Taq™ (TaKaRa) and the following thermal profile: 95°C for 3 minutes; 95°C for 10 seconds; 58°C for 30 seconds for 40 cycles.

    Techniques: Construct, Concentration Assay, Expressing, Quantitative RT-PCR

    ChIP assays. Capped mRNAs encoding mafK -FLAG, nrf2a -FLAG or bach1b -HA were microinjected into one-cell stage embryos, antibodies against FLAG or HA were used to pull down complexes of the protein with the DNA, and the DNAs were eluted. Specific primers and the eluted DNAs were used in PCR analyses to amplify the DNA fragments that contained cpa5 MARE sites, which were subsequently quantified by using qRT-PCR. (A) Electrophoresis analysis of the ChIP results showed that MafK bound to MARE-containing regulatory fragments of six zymogens. (B) Electrophoresis analysis of the cpa5 ChIP assay. Approximately 2.6-fold more Nrf2a than Bach1b protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in wild-type control larvae; whereas approximately 2.8-fold more Bach1b than Nrf2a protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in heme-deficient yquem/urod (−/−) larvae. Moreover, treatment of the heme-deficient yquem/urod (−/−) larvae with hemin reversed the situation so that approximately 1.2-fold more Nrf2a than Bach1b protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in hemin-treated yquem/urod (−/−) larvae. (C) qRT-PCR analysis of the cpa5 ChIP assay, the results of which were consistent with the gel electrophoresis analysis in B.

    Journal: Disease Models & Mechanisms

    Article Title: Heme acts through the Bach1b/Nrf2a-MafK pathway to regulate exocrine peptidase precursor genes in porphyric zebrafish

    doi: 10.1242/dmm.014951

    Figure Lengend Snippet: ChIP assays. Capped mRNAs encoding mafK -FLAG, nrf2a -FLAG or bach1b -HA were microinjected into one-cell stage embryos, antibodies against FLAG or HA were used to pull down complexes of the protein with the DNA, and the DNAs were eluted. Specific primers and the eluted DNAs were used in PCR analyses to amplify the DNA fragments that contained cpa5 MARE sites, which were subsequently quantified by using qRT-PCR. (A) Electrophoresis analysis of the ChIP results showed that MafK bound to MARE-containing regulatory fragments of six zymogens. (B) Electrophoresis analysis of the cpa5 ChIP assay. Approximately 2.6-fold more Nrf2a than Bach1b protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in wild-type control larvae; whereas approximately 2.8-fold more Bach1b than Nrf2a protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in heme-deficient yquem/urod (−/−) larvae. Moreover, treatment of the heme-deficient yquem/urod (−/−) larvae with hemin reversed the situation so that approximately 1.2-fold more Nrf2a than Bach1b protein (quantified with ImageJ) was associated with cpa5 promoters that harbored MARE sites in hemin-treated yquem/urod (−/−) larvae. (C) qRT-PCR analysis of the cpa5 ChIP assay, the results of which were consistent with the gel electrophoresis analysis in B.

    Article Snippet: RNA extraction and qRT-PCR Total RNA was extracted using the TRIzol® Reagent, according to the manufacturer’s instructions (Invitrogen). cDNA was synthesized by using reverse transcription with the M-MLV reverse transcription kit (Invitrogen), which was then used as the template for qRT-PCR analysis. qRT-PCR reactions were performed with the ABI StepOnePlus™ system, using SYBR® Premix Ex Taq™ (TaKaRa) and the following thermal profile: 95°C for 3 minutes; 95°C for 10 seconds; 58°C for 30 seconds for 40 cycles.

    Techniques: Chromatin Immunoprecipitation, Polymerase Chain Reaction, Quantitative RT-PCR, Electrophoresis, Nucleic Acid Electrophoresis

    The regulatory functions of BACH1 on exocrine zymogens are conserved, developmental delay of the exocrine pancreas in the HEP fish and a model for heme-mediated regulation of the exocrine peptidase precursor gene. (A) Overexpression of human BACH1 in zebrafish resulted in downregulation of the six peptidase precursor genes investigated. (B) Downregulation of exdpf ( Jiang et al., 2008 ), an exocrine pancreas marker, in zebrafish yquem/urod (−/−), as determined by qRT-PCR analysis, the results of which are consistent with those achieved by using in situ hybridization (shown in supplementary material Fig. S7A,B ). The primers used for the qRT-PCR analysis are listed in supplementary material Table S1 . Student’s t -tests were conducted. ** P

    Journal: Disease Models & Mechanisms

    Article Title: Heme acts through the Bach1b/Nrf2a-MafK pathway to regulate exocrine peptidase precursor genes in porphyric zebrafish

    doi: 10.1242/dmm.014951

    Figure Lengend Snippet: The regulatory functions of BACH1 on exocrine zymogens are conserved, developmental delay of the exocrine pancreas in the HEP fish and a model for heme-mediated regulation of the exocrine peptidase precursor gene. (A) Overexpression of human BACH1 in zebrafish resulted in downregulation of the six peptidase precursor genes investigated. (B) Downregulation of exdpf ( Jiang et al., 2008 ), an exocrine pancreas marker, in zebrafish yquem/urod (−/−), as determined by qRT-PCR analysis, the results of which are consistent with those achieved by using in situ hybridization (shown in supplementary material Fig. S7A,B ). The primers used for the qRT-PCR analysis are listed in supplementary material Table S1 . Student’s t -tests were conducted. ** P

    Article Snippet: RNA extraction and qRT-PCR Total RNA was extracted using the TRIzol® Reagent, according to the manufacturer’s instructions (Invitrogen). cDNA was synthesized by using reverse transcription with the M-MLV reverse transcription kit (Invitrogen), which was then used as the template for qRT-PCR analysis. qRT-PCR reactions were performed with the ABI StepOnePlus™ system, using SYBR® Premix Ex Taq™ (TaKaRa) and the following thermal profile: 95°C for 3 minutes; 95°C for 10 seconds; 58°C for 30 seconds for 40 cycles.

    Techniques: Fluorescence In Situ Hybridization, Over Expression, Marker, Quantitative RT-PCR, In Situ Hybridization

    Knockdown experiments with bach1b and nrf2a reveal their antagonism in the regulation of zymogen expression. (A) qRT-PCR analysis showed that bach1b knockdown results in the upregulation of the six peptidase precursor genes that we examined, whereas nrf2a knockdown resulted in their downregulation. Knockdown experiments were performed by microinjecting MOs that targeted bach1b or nrf2a into one-cell stage embryos. For bach1b knockdown experiments, an ATG MO and SPL MO were used. Reverse transcription PCR showed that the SPL MO effectively altered bach1b intron 2 splicing ( supplementary material Fig. S1A,B ). The ATG MO also effectively knocked down bach1b ( supplementary material Fig. S1C ). The results using the SPL MO are shown here. For nrf2a knockdown experiments, we used an ATG MO, of which the efficacy of knockdown has been confirmed previously ( Kobayashi et al., 2002 ; Kobayashi et al., 2009 ; Wang and Gallagher, 2013 ). The expression levels of the six peptidase precursor genes in the microinjected and control larvae at 84 hpf were determined by using qRT-PCR. (B) Representative images of in situ hybridization staining show that the upregulation of cpa5 resulted from bach1b knockdown and that its downregulation resulted from nrf2a knockdown, both results occurred specifically in the exocrine pancreas. Dorsal view, anterior to the left. (C) Mean optic densities of in situ hybridization staining of a group of larvae (10–12 each) corresponding to Fig. 4C were quantified by using ImageJ. (D) cpa5 morphant phenotypes. For individual larvae (84 hpf) of the bach1b knockdown overexpression group, the optic density values higher than the mean optic density value of its own group were marked as ‘strong’, and the optic density values lower than the mean optic density of the control group were marked as ‘weak’. For individual larvae of the nrf2a knockdown group, the optic density values lower than the mean optic density value of its own group were marked as ‘strong’, the optic density values higher than the mean optic density of the control group were marked as ‘weak’, and the values of optic density between ‘weak’ and ‘strong’ were marked as ‘medium’. The statistical significance of difference between means was determined by using one-way ANOVA and Tukey’s multiple comparison test ( n =9) with SPSS10.0.1. * P

    Journal: Disease Models & Mechanisms

    Article Title: Heme acts through the Bach1b/Nrf2a-MafK pathway to regulate exocrine peptidase precursor genes in porphyric zebrafish

    doi: 10.1242/dmm.014951

    Figure Lengend Snippet: Knockdown experiments with bach1b and nrf2a reveal their antagonism in the regulation of zymogen expression. (A) qRT-PCR analysis showed that bach1b knockdown results in the upregulation of the six peptidase precursor genes that we examined, whereas nrf2a knockdown resulted in their downregulation. Knockdown experiments were performed by microinjecting MOs that targeted bach1b or nrf2a into one-cell stage embryos. For bach1b knockdown experiments, an ATG MO and SPL MO were used. Reverse transcription PCR showed that the SPL MO effectively altered bach1b intron 2 splicing ( supplementary material Fig. S1A,B ). The ATG MO also effectively knocked down bach1b ( supplementary material Fig. S1C ). The results using the SPL MO are shown here. For nrf2a knockdown experiments, we used an ATG MO, of which the efficacy of knockdown has been confirmed previously ( Kobayashi et al., 2002 ; Kobayashi et al., 2009 ; Wang and Gallagher, 2013 ). The expression levels of the six peptidase precursor genes in the microinjected and control larvae at 84 hpf were determined by using qRT-PCR. (B) Representative images of in situ hybridization staining show that the upregulation of cpa5 resulted from bach1b knockdown and that its downregulation resulted from nrf2a knockdown, both results occurred specifically in the exocrine pancreas. Dorsal view, anterior to the left. (C) Mean optic densities of in situ hybridization staining of a group of larvae (10–12 each) corresponding to Fig. 4C were quantified by using ImageJ. (D) cpa5 morphant phenotypes. For individual larvae (84 hpf) of the bach1b knockdown overexpression group, the optic density values higher than the mean optic density value of its own group were marked as ‘strong’, and the optic density values lower than the mean optic density of the control group were marked as ‘weak’. For individual larvae of the nrf2a knockdown group, the optic density values lower than the mean optic density value of its own group were marked as ‘strong’, the optic density values higher than the mean optic density of the control group were marked as ‘weak’, and the values of optic density between ‘weak’ and ‘strong’ were marked as ‘medium’. The statistical significance of difference between means was determined by using one-way ANOVA and Tukey’s multiple comparison test ( n =9) with SPSS10.0.1. * P

    Article Snippet: RNA extraction and qRT-PCR Total RNA was extracted using the TRIzol® Reagent, according to the manufacturer’s instructions (Invitrogen). cDNA was synthesized by using reverse transcription with the M-MLV reverse transcription kit (Invitrogen), which was then used as the template for qRT-PCR analysis. qRT-PCR reactions were performed with the ABI StepOnePlus™ system, using SYBR® Premix Ex Taq™ (TaKaRa) and the following thermal profile: 95°C for 3 minutes; 95°C for 10 seconds; 58°C for 30 seconds for 40 cycles.

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, In Situ Hybridization, Staining, Over Expression

    Overexpression experiments with bach1b and nrf2a reveal their antagonism in the regulation of zymogen expression. (A) qRT-PCR analysis showed that bach1b overexpression resulted in the downregulation of six peptidase precursor genes – cpa5 , ctr1l , ctrb1 , ela2l , try and tryl – whereas nrf2a overexpression resulted in their upregulation. Overexpression experiments were performed by microinjecting bach1b or nrf2a capped mRNAs into one-cell stage embryos. The expression levels of the six peptidase precursor genes in the microinjected and control larvae at 84 hpf were determined by qRT-PCR analysis. (B) Representative images of in situ hybridization staining show that downregulation of cpa5 resulted from bach1b overexpression and that its upregulation resulted from nrf2a overexpression, both specifically in the exocrine pancreas. Dorsal view, anterior to the left. (C) Mean optic densities of in situ hybridization staining of a group of larvae (10–12 each) corresponding to Fig. 3B were quantified by using ImageJ. (D) cpa5 morphant phenotypes. For individual larvae (84 hpf) of the bach1b overexpression group, the optic density values lower than the mean optic density value of its own group were marked as ‘strong’, and the optic density values higher than the mean optic density of the control group were marked as ‘weak’. For individual larvae of the nrf2a overexpression group, the optic density values higher than the mean optic density value of its own group were marked as ‘strong’, the optic density values lower than the mean optic density of the control group were marked as ‘weak’, and the values of optic density between ‘weak’ and ‘strong’ marked as ‘medium’. The statistical significance of difference between means was determined by one-way ANOVA and Tukey’s multiple comparison test ( n =9) by using SPSS10.0.1. * P

    Journal: Disease Models & Mechanisms

    Article Title: Heme acts through the Bach1b/Nrf2a-MafK pathway to regulate exocrine peptidase precursor genes in porphyric zebrafish

    doi: 10.1242/dmm.014951

    Figure Lengend Snippet: Overexpression experiments with bach1b and nrf2a reveal their antagonism in the regulation of zymogen expression. (A) qRT-PCR analysis showed that bach1b overexpression resulted in the downregulation of six peptidase precursor genes – cpa5 , ctr1l , ctrb1 , ela2l , try and tryl – whereas nrf2a overexpression resulted in their upregulation. Overexpression experiments were performed by microinjecting bach1b or nrf2a capped mRNAs into one-cell stage embryos. The expression levels of the six peptidase precursor genes in the microinjected and control larvae at 84 hpf were determined by qRT-PCR analysis. (B) Representative images of in situ hybridization staining show that downregulation of cpa5 resulted from bach1b overexpression and that its upregulation resulted from nrf2a overexpression, both specifically in the exocrine pancreas. Dorsal view, anterior to the left. (C) Mean optic densities of in situ hybridization staining of a group of larvae (10–12 each) corresponding to Fig. 3B were quantified by using ImageJ. (D) cpa5 morphant phenotypes. For individual larvae (84 hpf) of the bach1b overexpression group, the optic density values lower than the mean optic density value of its own group were marked as ‘strong’, and the optic density values higher than the mean optic density of the control group were marked as ‘weak’. For individual larvae of the nrf2a overexpression group, the optic density values higher than the mean optic density value of its own group were marked as ‘strong’, the optic density values lower than the mean optic density of the control group were marked as ‘weak’, and the values of optic density between ‘weak’ and ‘strong’ marked as ‘medium’. The statistical significance of difference between means was determined by one-way ANOVA and Tukey’s multiple comparison test ( n =9) by using SPSS10.0.1. * P

    Article Snippet: RNA extraction and qRT-PCR Total RNA was extracted using the TRIzol® Reagent, according to the manufacturer’s instructions (Invitrogen). cDNA was synthesized by using reverse transcription with the M-MLV reverse transcription kit (Invitrogen), which was then used as the template for qRT-PCR analysis. qRT-PCR reactions were performed with the ABI StepOnePlus™ system, using SYBR® Premix Ex Taq™ (TaKaRa) and the following thermal profile: 95°C for 3 minutes; 95°C for 10 seconds; 58°C for 30 seconds for 40 cycles.

    Techniques: Over Expression, Expressing, Quantitative RT-PCR, In Situ Hybridization, Staining

    Survivin is induced in mature adipocytes from murine white adipose tissue by HFD feeding. Survivin expression in the subcutaneous and epididymal fat of 8-week-old mice was determined by qRT-PCR ( a ) and western blot ( b ); lung tumor tissues from nude mice used as a positive control. The SVF and adipocyte fraction (AF) were divided from the subcutaneous adipose tissue (SAT) and epididymal adipose tissue (EAT) of 8-week-old mice, then protein levels of Survivin in the SVF and adipocyte fraction divided from SAT ( c ) and EAT ( d ) were respectively determined. ( e – j ) C57BL/6 mice were fed with HFD or standard chow diet (CD) for 24 weeks; expression levels of Survivin in SAT and EAT were determined by qRT-PCR ( e ) and western blot ( f ). The SVF and adipocyte fractions were divided from SAT and EAT respectively, then the mRNA and protein content of Survivin in the SVF and adipocyte fraction divided from SAT ( g and h ) and EAT ( i and j ) was determined, respectively. ( k ) Microarray analysis of GSE27017, relative expression of Survivin in 6 months HFD and 6 months old ob/ob mice. * P

    Journal: Cell Death & Disease

    Article Title: Enhanced expression of Survivin has distinct roles in adipocyte homeostasis

    doi: 10.1038/cddis.2016.439

    Figure Lengend Snippet: Survivin is induced in mature adipocytes from murine white adipose tissue by HFD feeding. Survivin expression in the subcutaneous and epididymal fat of 8-week-old mice was determined by qRT-PCR ( a ) and western blot ( b ); lung tumor tissues from nude mice used as a positive control. The SVF and adipocyte fraction (AF) were divided from the subcutaneous adipose tissue (SAT) and epididymal adipose tissue (EAT) of 8-week-old mice, then protein levels of Survivin in the SVF and adipocyte fraction divided from SAT ( c ) and EAT ( d ) were respectively determined. ( e – j ) C57BL/6 mice were fed with HFD or standard chow diet (CD) for 24 weeks; expression levels of Survivin in SAT and EAT were determined by qRT-PCR ( e ) and western blot ( f ). The SVF and adipocyte fractions were divided from SAT and EAT respectively, then the mRNA and protein content of Survivin in the SVF and adipocyte fraction divided from SAT ( g and h ) and EAT ( i and j ) was determined, respectively. ( k ) Microarray analysis of GSE27017, relative expression of Survivin in 6 months HFD and 6 months old ob/ob mice. * P

    Article Snippet: Quantitative real-time PCR analysis Total RNA was isolated using the Trizol reagent (Invitrogen, 15596018, Carlsbad, CA, USA) and reverse transcription was performed using an iScript cDNA synthesis kit (Bio-Rad, 170-8891, Hercules, CA, USA) according to the manufacturer's instructions. qRT-PCR analysis analysis was carried out using SYBR Premix Ex Taq (Takara, RR820A, Japan) in a LightCycler480 PCR system (Roche, Germany).

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR, Western Blot, Positive Control, Microarray

    Survivin is expressed upon insulin exposure through the PI3K/mTOR pathway in mature adipocytes. During the differentiation process of 3T3-L1 cells and C3H10T1/2 cells, Survivin mRNA and protein levels were determined by qRT-PCR ( a and c ) and western blot ( b and d ). Survivin expression was detected by qRT-PCR and western blot in 3T3-L1 adipocytes, which were treated with 100 nM insulin for the indicated time ( e ) and insulin at the indicated concentrations for 24 h ( f ). Primary adipocytes, isolated and cultured from the SVF of subcutaneous fat, were induced to differentiate into mature adipocytes. Then Survivin content was determined by western blot after treating with 100 nM insulin for the indicated time ( g ) and insulin at the indicated concentrations for 24 h ( h ). Survivin content was immunobloted after rapamycin (10 nM) and LY294002 (50 μ M) for one hour prior to insulin exposure (100 nM) both in 3T3-L1 adipocytes and primary adipocytes ( i ). ( j ) 3T3-L1 adipocytes were incubated with insulin (100 nM) for 24 h, then incubated with or without HBSS for the indicated time. Immunoblots determined the levels of Survivin and GAPDH. ( k ) Protein levels of Survivin were detected by western blot in 3T3-L1 adipocyte overexpressing Survivin after incubation with HBSS for the indicated time. ( l ) The stability of ectopic expression of Survivin was evaluated in a cycloheximide (CHX, 100 μ g/ml) chasing experiment. Cells were harvested 0, 15, 30 or 60 min after the addition of CHX. The ectopic expression of Survivin protein level was evaluated using western blot. * P

    Journal: Cell Death & Disease

    Article Title: Enhanced expression of Survivin has distinct roles in adipocyte homeostasis

    doi: 10.1038/cddis.2016.439

    Figure Lengend Snippet: Survivin is expressed upon insulin exposure through the PI3K/mTOR pathway in mature adipocytes. During the differentiation process of 3T3-L1 cells and C3H10T1/2 cells, Survivin mRNA and protein levels were determined by qRT-PCR ( a and c ) and western blot ( b and d ). Survivin expression was detected by qRT-PCR and western blot in 3T3-L1 adipocytes, which were treated with 100 nM insulin for the indicated time ( e ) and insulin at the indicated concentrations for 24 h ( f ). Primary adipocytes, isolated and cultured from the SVF of subcutaneous fat, were induced to differentiate into mature adipocytes. Then Survivin content was determined by western blot after treating with 100 nM insulin for the indicated time ( g ) and insulin at the indicated concentrations for 24 h ( h ). Survivin content was immunobloted after rapamycin (10 nM) and LY294002 (50 μ M) for one hour prior to insulin exposure (100 nM) both in 3T3-L1 adipocytes and primary adipocytes ( i ). ( j ) 3T3-L1 adipocytes were incubated with insulin (100 nM) for 24 h, then incubated with or without HBSS for the indicated time. Immunoblots determined the levels of Survivin and GAPDH. ( k ) Protein levels of Survivin were detected by western blot in 3T3-L1 adipocyte overexpressing Survivin after incubation with HBSS for the indicated time. ( l ) The stability of ectopic expression of Survivin was evaluated in a cycloheximide (CHX, 100 μ g/ml) chasing experiment. Cells were harvested 0, 15, 30 or 60 min after the addition of CHX. The ectopic expression of Survivin protein level was evaluated using western blot. * P

    Article Snippet: Quantitative real-time PCR analysis Total RNA was isolated using the Trizol reagent (Invitrogen, 15596018, Carlsbad, CA, USA) and reverse transcription was performed using an iScript cDNA synthesis kit (Bio-Rad, 170-8891, Hercules, CA, USA) according to the manufacturer's instructions. qRT-PCR analysis analysis was carried out using SYBR Premix Ex Taq (Takara, RR820A, Japan) in a LightCycler480 PCR system (Roche, Germany).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Isolation, Cell Culture, Incubation

    Quantitative analysis of lncRNA CRNDE-h using qRT-PCR in 305 colorectal disease tissues. Notes: ( A ) The expression level of lncRNA CRNDE-h was significantly higher in the CRC and Ad group compared with the NC, IBD, and HP group, and its expression was normalized by GAPDH in each sample (*** P

    Journal: OncoTargets and therapy

    Article Title: Increased expression of the long noncoding RNA CRNDE-h indicates a poor prognosis in colorectal cancer, and is positively correlated with IRX5 mRNA expression

    doi: 10.2147/OTT.S98268

    Figure Lengend Snippet: Quantitative analysis of lncRNA CRNDE-h using qRT-PCR in 305 colorectal disease tissues. Notes: ( A ) The expression level of lncRNA CRNDE-h was significantly higher in the CRC and Ad group compared with the NC, IBD, and HP group, and its expression was normalized by GAPDH in each sample (*** P

    Article Snippet: qRT-PCR qRT-PCR analysis was performed using Power SYBR Green (TaKaRa) on a CFX96 Real-Time PCR Detection System, which was also used for data collection.

    Techniques: Quantitative RT-PCR, Expressing

    Relative miR-30a expression and colony survival analysis of A549 and H460 cells. (A and B) Relative miR-30a expression in (A) A549 and (B) H460 cells after transfected with hsa-miR-30a agomir, hsa-miR-30a antagomir and their negative controls, detected by qRT-PCR (**P

    Journal: Oncology Reports

    Article Title: miR-30a radiosensitizes non-small cell lung cancer by targeting ATF1 that is involved in the phosphorylation of ATM

    doi: 10.3892/or.2017.5448

    Figure Lengend Snippet: Relative miR-30a expression and colony survival analysis of A549 and H460 cells. (A and B) Relative miR-30a expression in (A) A549 and (B) H460 cells after transfected with hsa-miR-30a agomir, hsa-miR-30a antagomir and their negative controls, detected by qRT-PCR (**P

    Article Snippet: RNA extraction and qRT-PCR analysis RNA extraction kit (Takara Bio, Inc., Shiga, China) was used to isolate total RNA and TRIzol (Invitrogen Life Technologies) to extract microRNA, following the manufacturer's instructions.

    Techniques: Expressing, Transfection, Quantitative RT-PCR

    miR-30a directly targets the 3′UTR of ATF1. (A) The putative targeted sites of ATF1 in 3′UTR. (B) Schematic diagram of miR-30a targeting the 3′UTR of ATF1. (C) Relative luciferase activity in co-transfected groups of miR-30a agomir and pmirGLO vector, miR-30a agomir and pmirGLO ATF1-wild, miR-30a agomir and pmirGLO ATF1-mutant. (D) Relative ATF1 mRNA expression detected by qRT-PCR (**P

    Journal: Oncology Reports

    Article Title: miR-30a radiosensitizes non-small cell lung cancer by targeting ATF1 that is involved in the phosphorylation of ATM

    doi: 10.3892/or.2017.5448

    Figure Lengend Snippet: miR-30a directly targets the 3′UTR of ATF1. (A) The putative targeted sites of ATF1 in 3′UTR. (B) Schematic diagram of miR-30a targeting the 3′UTR of ATF1. (C) Relative luciferase activity in co-transfected groups of miR-30a agomir and pmirGLO vector, miR-30a agomir and pmirGLO ATF1-wild, miR-30a agomir and pmirGLO ATF1-mutant. (D) Relative ATF1 mRNA expression detected by qRT-PCR (**P

    Article Snippet: RNA extraction and qRT-PCR analysis RNA extraction kit (Takara Bio, Inc., Shiga, China) was used to isolate total RNA and TRIzol (Invitrogen Life Technologies) to extract microRNA, following the manufacturer's instructions.

    Techniques: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Expressing, Quantitative RT-PCR

    Expression levels of ZmJAZ14 in different maize organs. The relative expression level of ZmJAZ14 was normalized with ZmActin1 . Data from qRT-PCR were analyzed according to the 2 -ΔΔCt method. The error bars indicate standard deviations.

    Journal: PLoS ONE

    Article Title: A Maize Jasmonate Zim-Domain Protein, ZmJAZ14, Associates with the JA, ABA, and GA Signaling Pathways in Transgenic Arabidopsis

    doi: 10.1371/journal.pone.0121824

    Figure Lengend Snippet: Expression levels of ZmJAZ14 in different maize organs. The relative expression level of ZmJAZ14 was normalized with ZmActin1 . Data from qRT-PCR were analyzed according to the 2 -ΔΔCt method. The error bars indicate standard deviations.

    Article Snippet: qRT-PCR analysis RNA was extracted using Trizol reagent (TaKaRa, Otsu, Japan) following the manufacturer’s instructions and then treated with RNase-free DNase I (Promega, Madison, WI) to remove genomic DNA.

    Techniques: Expressing, Quantitative RT-PCR

    miR-140 Expression Is Reversely Correlated with CRC Development and Metastasis, Whereas Smad3 Expression Is Positively Correlated with CRC Development (A) A cohort of CRC specimens, including 31 paired CRC tissues and adjacent colorectal mucosa, were selected, and the expression levels of miR-140 were measured by real-time qRT-PCR. MiR-140 was significantly downregulated in the primary CRC tissues compared to the adjacent normal mucosa. (B) Then, 13 out of 31 cases of CRC patients with lymph node or liver metastasis were selected, and RNA was extracted from the metastatic tumor tissues. miR-140 was significantly downregulated in the lymph node, and liver metastatic tumors were compared with the paired primary tumor tissues. The expression level of miR-140 was normalized by the internal control RNU6B in each sample. (C) Immunohistochemistry analysis of Smad3 expression was performed in the CRC cohort. Smad3 protein was overexpressed in the CRC tissues compared to the adjacent normal mucosa. Each experiment was repeated 3 times; data are presented as means ± SD. *p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: MicroRNA-140 Inhibits the Epithelial-Mesenchymal Transition and Metastasis in Colorectal Cancer

    doi: 10.1016/j.omtn.2017.12.022

    Figure Lengend Snippet: miR-140 Expression Is Reversely Correlated with CRC Development and Metastasis, Whereas Smad3 Expression Is Positively Correlated with CRC Development (A) A cohort of CRC specimens, including 31 paired CRC tissues and adjacent colorectal mucosa, were selected, and the expression levels of miR-140 were measured by real-time qRT-PCR. MiR-140 was significantly downregulated in the primary CRC tissues compared to the adjacent normal mucosa. (B) Then, 13 out of 31 cases of CRC patients with lymph node or liver metastasis were selected, and RNA was extracted from the metastatic tumor tissues. miR-140 was significantly downregulated in the lymph node, and liver metastatic tumors were compared with the paired primary tumor tissues. The expression level of miR-140 was normalized by the internal control RNU6B in each sample. (C) Immunohistochemistry analysis of Smad3 expression was performed in the CRC cohort. Smad3 protein was overexpressed in the CRC tissues compared to the adjacent normal mucosa. Each experiment was repeated 3 times; data are presented as means ± SD. *p

    Article Snippet: For real-time qRT-PCR analysis of mRNA expression, cDNA was synthesized using the PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Dalian, China).

    Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

    qRT-PCR confirmation of sequencing results in chickens with or without ALV-J infection. (A) For mRNA. (B) For lncRNA.

    Journal: Frontiers in Physiology

    Article Title: Comprehensive Transcriptome Analysis Reveals Competing Endogenous RNA Networks During Avian Leukosis Virus, Subgroup J-Induced Tumorigenesis in Chickens

    doi: 10.3389/fphys.2018.00996

    Figure Lengend Snippet: qRT-PCR confirmation of sequencing results in chickens with or without ALV-J infection. (A) For mRNA. (B) For lncRNA.

    Article Snippet: qRT-Time PCR Analysis qRT-PCR analysis was performed to validate the sequencing results and identify core transcripts related to ALV-J-induced tumorigenesis. qRT-PCR analysis was conducted using SYBR Premix Ex TaqTM II (TaKaRa, Shiga, Japan) and an QuantStudio 5 real-time PCR instrument after cDNA synthesis with the PrimeScriptTM RT Reagent Kit with gDNA Eraser (TaKaRa).

    Techniques: Quantitative RT-PCR, Sequencing, Infection

    Phenotypic analysis of transgenic tobacco overexpressing CsMYB6A and AtPAP1 . ( a ) Leaves of transgenic plants ( CsMYB6A and AtPAP1 ) in comparion with empty-vector transgenic plants; ( b ) Total concentrations of anthocyanins and flavonols of transgenic plants. The relative flavonol concentration was calculated as the ratio between the total peak area at 350 nm; ( c ) The relative gene expression involved in flavonoid biosynthesis in transgemic plants compared with the empty vector. Relative FPKM and expression data were obtained from RNA-sequencing and qRT-PCR analysis, respectively.

    Journal: Scientific Reports

    Article Title: Isolation and Characterization of Key Genes that Promote Flavonoid Accumulation in Purple-leaf Tea (Camellia sinensis L.)

    doi: 10.1038/s41598-017-18133-z

    Figure Lengend Snippet: Phenotypic analysis of transgenic tobacco overexpressing CsMYB6A and AtPAP1 . ( a ) Leaves of transgenic plants ( CsMYB6A and AtPAP1 ) in comparion with empty-vector transgenic plants; ( b ) Total concentrations of anthocyanins and flavonols of transgenic plants. The relative flavonol concentration was calculated as the ratio between the total peak area at 350 nm; ( c ) The relative gene expression involved in flavonoid biosynthesis in transgemic plants compared with the empty vector. Relative FPKM and expression data were obtained from RNA-sequencing and qRT-PCR analysis, respectively.

    Article Snippet: qRT-PCR analysis Quantitative real-time PCR (qRT-PCR) was performed using the SYBR® Premix Ex Taq™ II (Perfect Real Time) kit (Takara, Japan) on a Peltier Thermal Cycler PTC200 (Bio-Rad, USA) with gene-specific primer pairs (Table ).

    Techniques: Transgenic Assay, Plasmid Preparation, Concentration Assay, Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    The expression levels of 6 lignin pathway genes (C3H, C4H, CAD, CCoA-OMT, CCR and F5H) and Myb46 and NAC genes using qRT-PCR. Blue lines mean the hard-endocarp hawthorn (H8), red lines mean the soft-endocarp hawthorn (S7). Y-axis represents normalized fold expression, X-axis is DAB (Days after bloom).

    Journal: PLoS ONE

    Article Title: Transcript Assembly and Quantification by RNA-Seq Reveals Differentially Expressed Genes between Soft-Endocarp and Hard-Endocarp Hawthorns

    doi: 10.1371/journal.pone.0072910

    Figure Lengend Snippet: The expression levels of 6 lignin pathway genes (C3H, C4H, CAD, CCoA-OMT, CCR and F5H) and Myb46 and NAC genes using qRT-PCR. Blue lines mean the hard-endocarp hawthorn (H8), red lines mean the soft-endocarp hawthorn (S7). Y-axis represents normalized fold expression, X-axis is DAB (Days after bloom).

    Article Snippet: Quantitative RT-PCR (qRT-PCR) Analysis The cDNA was synthesized from total RNA using Reverse Transcriptase XL (AMV) (TaKaRa, Japan) in a 20 µL reaction system.

    Techniques: Expressing, Quantitative RT-PCR

    Analysis of flavonoid contents and expression level of anthocyanin biosynthesis-related genes during eight time points from 8:00 am to 5:00 pm in young and mature leaves of the Malus crabapple ever-red cultivar ‘Royalty.’ (A) The content of the main flavonoid compounds at eight time points in young and mature leaves of ‘Royalty.’ (B,C) The expression patterns of McCHS , McF3H , McF3’H , McDFR , McANS , McUFGT , McMYB10 , McCOP1-1 , and McCOP1-2 were analyzed in the young and mature leaves of ‘Royalty’ by qRT-PCR. Malus 18 S (DQ341382) was used as the reference gene. Error bars indicate the standard error of the mean ± SE of three replicate measurements. The expression levels of flavonoid-related genes were calculated using CFX-Manager-3-1 software (Bio-Rad). Different letters above the bars indicate significantly different values ( P

    Journal: Frontiers in Plant Science

    Article Title: McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    doi: 10.3389/fpls.2018.00704

    Figure Lengend Snippet: Analysis of flavonoid contents and expression level of anthocyanin biosynthesis-related genes during eight time points from 8:00 am to 5:00 pm in young and mature leaves of the Malus crabapple ever-red cultivar ‘Royalty.’ (A) The content of the main flavonoid compounds at eight time points in young and mature leaves of ‘Royalty.’ (B,C) The expression patterns of McCHS , McF3H , McF3’H , McDFR , McANS , McUFGT , McMYB10 , McCOP1-1 , and McCOP1-2 were analyzed in the young and mature leaves of ‘Royalty’ by qRT-PCR. Malus 18 S (DQ341382) was used as the reference gene. Error bars indicate the standard error of the mean ± SE of three replicate measurements. The expression levels of flavonoid-related genes were calculated using CFX-Manager-3-1 software (Bio-Rad). Different letters above the bars indicate significantly different values ( P

    Article Snippet: Quantitative RT-PCR Analysis (qRT-PCR) The qRT-PCR experiment was performed using the SYBR® Premix Ex TaqTM II (Perfect Real Time) (TaKaRa, Ohtsu, Japan) and the CFX96TM Real Time System (Bio-Rad, Hercules, CA, United States).

    Techniques: Expressing, Quantitative RT-PCR, Software

    Silencing of McMYB10 in the ever-red cultivar ‘Royalty’. (A) After 14 days infiltration, the faded red leaf phenotype was observed. (B) Flavonoid contents in infiltrated ‘Royalty’ plantlets in μg/g fresh weight (FW). (C) qRT-PCR was conducted to analysis the expression levels of anthocyanin biosynthesis-related genes. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Journal: Frontiers in Plant Science

    Article Title: McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    doi: 10.3389/fpls.2018.00704

    Figure Lengend Snippet: Silencing of McMYB10 in the ever-red cultivar ‘Royalty’. (A) After 14 days infiltration, the faded red leaf phenotype was observed. (B) Flavonoid contents in infiltrated ‘Royalty’ plantlets in μg/g fresh weight (FW). (C) qRT-PCR was conducted to analysis the expression levels of anthocyanin biosynthesis-related genes. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Article Snippet: Quantitative RT-PCR Analysis (qRT-PCR) The qRT-PCR experiment was performed using the SYBR® Premix Ex TaqTM II (Perfect Real Time) (TaKaRa, Ohtsu, Japan) and the CFX96TM Real Time System (Bio-Rad, Hercules, CA, United States).

    Techniques: Quantitative RT-PCR, Expressing

    Different light treatments in crabapple foliage. (A) ‘Flame’ tissue culture plants under different light treatments. After 3 days treatment, red color appeared in the plants treated with light for a 24-h period (right panel). (B) High pressure liquid chromatography (HPLC) analysis showing the variation of flavonoids in crabapple leaves under different light treatments. (C) qRT-PCR was conducted to analysis the expression levels of anthocyanin biosynthesis-related genes in ‘Flame’ plantlets. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Journal: Frontiers in Plant Science

    Article Title: McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    doi: 10.3389/fpls.2018.00704

    Figure Lengend Snippet: Different light treatments in crabapple foliage. (A) ‘Flame’ tissue culture plants under different light treatments. After 3 days treatment, red color appeared in the plants treated with light for a 24-h period (right panel). (B) High pressure liquid chromatography (HPLC) analysis showing the variation of flavonoids in crabapple leaves under different light treatments. (C) qRT-PCR was conducted to analysis the expression levels of anthocyanin biosynthesis-related genes in ‘Flame’ plantlets. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Article Snippet: Quantitative RT-PCR Analysis (qRT-PCR) The qRT-PCR experiment was performed using the SYBR® Premix Ex TaqTM II (Perfect Real Time) (TaKaRa, Ohtsu, Japan) and the CFX96TM Real Time System (Bio-Rad, Hercules, CA, United States).

    Techniques: High Performance Liquid Chromatography, Quantitative RT-PCR, Expressing

    Transiently silencing of MYB10 in apple fruit. MYB10 was silenced using the TRV-GFP- MYB10 vector. Apple fruit vacuum infiltration with the empty TRV vectors were used as controls. (A) Apple fruit peel coloration around the infiltrated sites of suppressed (TRV-GFP- MYB10 ) apples. (B) Flavonoid compound levels around the infiltrated sites of apple peels were analyzed by HPLC in μg/g FW. (C) qRT-PCR was conducted to analyze the expression levels around the infiltrated sites, the expression level of MYB10 represent the transcription level of endogenous MdMYB10 in silenced fruits. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Journal: Frontiers in Plant Science

    Article Title: McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    doi: 10.3389/fpls.2018.00704

    Figure Lengend Snippet: Transiently silencing of MYB10 in apple fruit. MYB10 was silenced using the TRV-GFP- MYB10 vector. Apple fruit vacuum infiltration with the empty TRV vectors were used as controls. (A) Apple fruit peel coloration around the infiltrated sites of suppressed (TRV-GFP- MYB10 ) apples. (B) Flavonoid compound levels around the infiltrated sites of apple peels were analyzed by HPLC in μg/g FW. (C) qRT-PCR was conducted to analyze the expression levels around the infiltrated sites, the expression level of MYB10 represent the transcription level of endogenous MdMYB10 in silenced fruits. Error bars indicate the standard error of the mean ± SE of three replicate measurements. Different letters above the bars indicate significantly different values ( P

    Article Snippet: Quantitative RT-PCR Analysis (qRT-PCR) The qRT-PCR experiment was performed using the SYBR® Premix Ex TaqTM II (Perfect Real Time) (TaKaRa, Ohtsu, Japan) and the CFX96TM Real Time System (Bio-Rad, Hercules, CA, United States).

    Techniques: Plasmid Preparation, High Performance Liquid Chromatography, Quantitative RT-PCR, Expressing

    Analysis of flavonoid contents and expression level of anthocyanin biosynthesis-related genes during five leaf developmental stages in the ever-red crabapple cultivar ‘Royalty’ and ever-green crabapple cultivar ‘Flame’. (A) The phenotypes of leaves in five leaf developmental stages. (B) Flavonoid contents in five leaf developmental stages of ‘Royalty’ and ‘Flame.’ (C) The expression patterns of McCHS , McF3H , McF3’H , McDFR , McANS , McUFGT , McMYB10 , McCOP1-1 , and McCOP1-2 were analyzed by qRT-PCR in the leaves of ‘Royalty’ and ‘Flame.’ Malus 18 S (DQ341382) as the reference gene. S1 to S5 represents 3 days after budding, 9 days after budding, 15 days after budding, 21 days after budding, 30 days after budding. Error bars indicate the standard error of the mean ± SE of three replicate measurements. The expression levels of flavonoid-related genes were calculated using CFX-Manager-3-1 software (Bio-Rad). Scale bars = 1 cm. Different letters above the bars indicate significantly different values ( P

    Journal: Frontiers in Plant Science

    Article Title: McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    doi: 10.3389/fpls.2018.00704

    Figure Lengend Snippet: Analysis of flavonoid contents and expression level of anthocyanin biosynthesis-related genes during five leaf developmental stages in the ever-red crabapple cultivar ‘Royalty’ and ever-green crabapple cultivar ‘Flame’. (A) The phenotypes of leaves in five leaf developmental stages. (B) Flavonoid contents in five leaf developmental stages of ‘Royalty’ and ‘Flame.’ (C) The expression patterns of McCHS , McF3H , McF3’H , McDFR , McANS , McUFGT , McMYB10 , McCOP1-1 , and McCOP1-2 were analyzed by qRT-PCR in the leaves of ‘Royalty’ and ‘Flame.’ Malus 18 S (DQ341382) as the reference gene. S1 to S5 represents 3 days after budding, 9 days after budding, 15 days after budding, 21 days after budding, 30 days after budding. Error bars indicate the standard error of the mean ± SE of three replicate measurements. The expression levels of flavonoid-related genes were calculated using CFX-Manager-3-1 software (Bio-Rad). Scale bars = 1 cm. Different letters above the bars indicate significantly different values ( P

    Article Snippet: Quantitative RT-PCR Analysis (qRT-PCR) The qRT-PCR experiment was performed using the SYBR® Premix Ex TaqTM II (Perfect Real Time) (TaKaRa, Ohtsu, Japan) and the CFX96TM Real Time System (Bio-Rad, Hercules, CA, United States).

    Techniques: Expressing, Quantitative RT-PCR, Software

    PAI‐1 upregulated the expression of CCL5 in SKBR‐3/SI cells. A, qRT‐PCR verified the increased expression of CCL5 in SKBR‐3/SI cells. B,C, The expression level of CCL5 and the secretion level of CCL5 were detected in SKBR‐3 after treated with 10 ng/mL of rPAI‐1and PAI‐039 for 48 h by qRT‐PCR and ELISA, respectively. D, qRT‐PCR analysis of PAI‐1 mRNA expression post‐siRNA knockdown of PAI‐1 in SKBR‐3/SI cells. E, PAI‐1 and CCL5 expression were determined in SKBR‐3/SI after knockdown of PAI‐1 by western blot. The error bars represent the mean of 3 separate determinations ± SD. * P

    Journal: Cancer Science

    Article Title: PAI‐1 induces Src inhibitor resistance via CCL5 in HER2‐positive breast cancer cells. PAI‐1 induces Src inhibitor resistance via CCL5 in HER2‐positive breast cancer cells

    doi: 10.1111/cas.13593

    Figure Lengend Snippet: PAI‐1 upregulated the expression of CCL5 in SKBR‐3/SI cells. A, qRT‐PCR verified the increased expression of CCL5 in SKBR‐3/SI cells. B,C, The expression level of CCL5 and the secretion level of CCL5 were detected in SKBR‐3 after treated with 10 ng/mL of rPAI‐1and PAI‐039 for 48 h by qRT‐PCR and ELISA, respectively. D, qRT‐PCR analysis of PAI‐1 mRNA expression post‐siRNA knockdown of PAI‐1 in SKBR‐3/SI cells. E, PAI‐1 and CCL5 expression were determined in SKBR‐3/SI after knockdown of PAI‐1 by western blot. The error bars represent the mean of 3 separate determinations ± SD. * P

    Article Snippet: For quantitative real‐time PCR (qRT‐PCR) analysis of PAI‐1, CCL5 and CCR5 mRNA expression, cDNA obtained by reverse transcription of 2 μg total RNA was synthesized using a PrimeScript RT Master Mix (Perfect Real Time) Kit (RR036A, Takara, China), followed by PCR using Power SYBR Green PCR Master Mix (Life Technology, USA). β‐actin was used as an internal control.

    Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot

    MJ inhibits cell proliferation through BCRC-3/miR-182-5p/p27 axis in BC cells. a Western blot analysis of the expression levels of cell cycle related proteins (p27, CyclinE, p21) in EJ or T24 T cells at different time points after MJ treatment. b Western blot analysis of the expression levels of p27 after MJ treatment in cells that were stably transfected with shP27 cells. c d Cell cycle distributions in p27 knockdown cells after MJ treatment were presented by flow cytometry. e qRT-PCR analysis of the expression levels of p27 in EJ or T24 T cells after MJ treatment for different time points. f g The luciferase activities of p27 3’UTR and promoter in EJ or T24 T cells after MJ treatment for 24 h. h i qRT-PCR analysis of the expression levels of BCRC-3 ( h ) and miR-182-5p ( i ) in EJ or T24 T cells after MJ treatment for different time points. j k Western blot analysis of the expression levels of p27 after MJ treatment in EJ or T24 T cells transfected with miR-182-5p mimics or si BCRC-3. (The data indicated mean ± SEM of three experiments. Student’s t -test analyzed the difference in d - i . P

    Journal: Molecular Cancer

    Article Title: Circular RNA BCRC-3 suppresses bladder cancer proliferation through miR-182-5p/p27 axis

    doi: 10.1186/s12943-018-0892-z

    Figure Lengend Snippet: MJ inhibits cell proliferation through BCRC-3/miR-182-5p/p27 axis in BC cells. a Western blot analysis of the expression levels of cell cycle related proteins (p27, CyclinE, p21) in EJ or T24 T cells at different time points after MJ treatment. b Western blot analysis of the expression levels of p27 after MJ treatment in cells that were stably transfected with shP27 cells. c d Cell cycle distributions in p27 knockdown cells after MJ treatment were presented by flow cytometry. e qRT-PCR analysis of the expression levels of p27 in EJ or T24 T cells after MJ treatment for different time points. f g The luciferase activities of p27 3’UTR and promoter in EJ or T24 T cells after MJ treatment for 24 h. h i qRT-PCR analysis of the expression levels of BCRC-3 ( h ) and miR-182-5p ( i ) in EJ or T24 T cells after MJ treatment for different time points. j k Western blot analysis of the expression levels of p27 after MJ treatment in EJ or T24 T cells transfected with miR-182-5p mimics or si BCRC-3. (The data indicated mean ± SEM of three experiments. Student’s t -test analyzed the difference in d - i . P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was carried out using the SYBR Premix Ex TaqTM kit (Takara).

    Techniques: Western Blot, Expressing, Stable Transfection, Transfection, Flow Cytometry, Cytometry, Quantitative RT-PCR, Luciferase

    The expression of BCRC-3 in cell lines and tissues, and the subcellular location of BCRC-3. a qRT-PCR assay with divergent primers indicated the low expression of BCRC-3 in 47 pairs of human BC compared with their adjacent normal tissues. b The expression of BCRC-3 in SV-HUC-1, EJ and T24 T cell lines were measured by qRT-PCR. c Schematic diagram demonstrated that nine exons derived from PSMD1 constituted BCRC-3. The existence of BCRC-3 was proved by RT–PCR and its back splicing junction was verified by Sanger sequencing. Red arrow indicated the special splicing junction of BCRC-3. d RT-PCR assay with divergent or convergent primers indicating the existence of BCRC-3 in SV-HUC-1, EJ, T24 T cell lines and three BC tissues. GAPDH was used as negative control. e qRT-PCR analysis of the expression of BCRC-3 after RNase R treatmet in EJ or T24 T cells. f RNA-FISH indicated the location of BCRC-3 in T24 T cells. U6 was used as negative control and 18S was used as positive control. Nuclei were stained blue by DAPI. BCRC-3, U6 and 18S were stained red with cy3 (scale bar, 10 μm). (Data are mean ± SEM of three experiments. Student’s t -test analyzed the difference in a - b , e . * P

    Journal: Molecular Cancer

    Article Title: Circular RNA BCRC-3 suppresses bladder cancer proliferation through miR-182-5p/p27 axis

    doi: 10.1186/s12943-018-0892-z

    Figure Lengend Snippet: The expression of BCRC-3 in cell lines and tissues, and the subcellular location of BCRC-3. a qRT-PCR assay with divergent primers indicated the low expression of BCRC-3 in 47 pairs of human BC compared with their adjacent normal tissues. b The expression of BCRC-3 in SV-HUC-1, EJ and T24 T cell lines were measured by qRT-PCR. c Schematic diagram demonstrated that nine exons derived from PSMD1 constituted BCRC-3. The existence of BCRC-3 was proved by RT–PCR and its back splicing junction was verified by Sanger sequencing. Red arrow indicated the special splicing junction of BCRC-3. d RT-PCR assay with divergent or convergent primers indicating the existence of BCRC-3 in SV-HUC-1, EJ, T24 T cell lines and three BC tissues. GAPDH was used as negative control. e qRT-PCR analysis of the expression of BCRC-3 after RNase R treatmet in EJ or T24 T cells. f RNA-FISH indicated the location of BCRC-3 in T24 T cells. U6 was used as negative control and 18S was used as positive control. Nuclei were stained blue by DAPI. BCRC-3, U6 and 18S were stained red with cy3 (scale bar, 10 μm). (Data are mean ± SEM of three experiments. Student’s t -test analyzed the difference in a - b , e . * P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was carried out using the SYBR Premix Ex TaqTM kit (Takara).

    Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Sequencing, Negative Control, Fluorescence In Situ Hybridization, Positive Control, Staining

    Enforced expression of BCRC-3 inhibits cell proliferation in vitro and impairs tumor growth in vivo. a qRT-PCR analysis verified the effective overexpression of BCRC-3 after transfection of the vector for 48 h in EJ or T24 T cells. b Cell cycle distributions in BCRC-3 overexpression cells were presented by flow cytometry. c Cloning formation assay of EJ or T24 T cells transfected with BCRC-3 overexpression vectors. Colonies with more than 50 cells were counted. d e Observation of DNA synthesis of EJ or T24 T cells transfected with BCRC-3 overexpression vectors by EdU assay. f T24 T cells with stable overexpression of BCRC-3 or vector were injected in the flanks (5 × 10 6 cells per mouse, n = 6 for each group). Tumors collected from mice were measured and weighed after one month of hypodermic injection. (Data are mean ± SEM of three experiments. Student’s t -test analyzed the difference in a - c , e - f . * P

    Journal: Molecular Cancer

    Article Title: Circular RNA BCRC-3 suppresses bladder cancer proliferation through miR-182-5p/p27 axis

    doi: 10.1186/s12943-018-0892-z

    Figure Lengend Snippet: Enforced expression of BCRC-3 inhibits cell proliferation in vitro and impairs tumor growth in vivo. a qRT-PCR analysis verified the effective overexpression of BCRC-3 after transfection of the vector for 48 h in EJ or T24 T cells. b Cell cycle distributions in BCRC-3 overexpression cells were presented by flow cytometry. c Cloning formation assay of EJ or T24 T cells transfected with BCRC-3 overexpression vectors. Colonies with more than 50 cells were counted. d e Observation of DNA synthesis of EJ or T24 T cells transfected with BCRC-3 overexpression vectors by EdU assay. f T24 T cells with stable overexpression of BCRC-3 or vector were injected in the flanks (5 × 10 6 cells per mouse, n = 6 for each group). Tumors collected from mice were measured and weighed after one month of hypodermic injection. (Data are mean ± SEM of three experiments. Student’s t -test analyzed the difference in a - c , e - f . * P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was carried out using the SYBR Premix Ex TaqTM kit (Takara).

    Techniques: Expressing, In Vitro, In Vivo, Quantitative RT-PCR, Over Expression, Transfection, Plasmid Preparation, Flow Cytometry, Cytometry, Clone Assay, Tube Formation Assay, DNA Synthesis, EdU Assay, Injection, Mouse Assay

    BCRC-3 sponges miR-182-5p, and overexpression of BCRC-3 reverses the down-regulation effect of miR-182-5p on p27. a The luciferase activity of p27 3’UTR and promoter after transfection with BCRC-3 overexpression vectors in EJ or T24 T cell lines. b The relative luciferase activities of p27 3’UTR and truncates in EJ or T24 T cells transfected with BCRC-3 overexpression vector. Schematic Sequence of p27 3’UTR full length and truncates (up panel). c d The efficiency of BCRC-3 pull-down assay was verified by RT-PCR ( c ) or real-time PCR ( d ). GAPDH was used as negative control. Relative level of BCRC-3 was normalized to input. e qRT-PCR analysis of the expression of 7 candidates in the EJ and T24 T lysates after biotin-BCRC-3 pull-down assay. f RNA-FISH detection of BCRC-3 in T24 T cells. Nuclei were stained blue with DAPI. BCRC-3 was stained red with cy3. Locked nucleic acid miR-182-5p probes were labeled with Dig (scale bar, 10 μm). g qRT-PCR analysis of the expression levels of BCRC-3 or GAPDH in the EJ and T24 T lysates after biotin-miR-182-5p pull-down assay. GAPDH was used as negative control. h The luciferase activities of wild type p27 3’UTR and mutant p27 3’UTR after transfection with miR-182-5p mimic in EJ or T24 T cell lines. i j qRT-PCR and western blot analysis of the expression levels of p27 in EJ or T24 T cells after co-transfection with BCRC-3 overexpression vector and miR-182-5p mimics. (Data are mean ± SEM of three experiments. Student’s t- test analyzed the difference in a - b , d - e , g - i . * P

    Journal: Molecular Cancer

    Article Title: Circular RNA BCRC-3 suppresses bladder cancer proliferation through miR-182-5p/p27 axis

    doi: 10.1186/s12943-018-0892-z

    Figure Lengend Snippet: BCRC-3 sponges miR-182-5p, and overexpression of BCRC-3 reverses the down-regulation effect of miR-182-5p on p27. a The luciferase activity of p27 3’UTR and promoter after transfection with BCRC-3 overexpression vectors in EJ or T24 T cell lines. b The relative luciferase activities of p27 3’UTR and truncates in EJ or T24 T cells transfected with BCRC-3 overexpression vector. Schematic Sequence of p27 3’UTR full length and truncates (up panel). c d The efficiency of BCRC-3 pull-down assay was verified by RT-PCR ( c ) or real-time PCR ( d ). GAPDH was used as negative control. Relative level of BCRC-3 was normalized to input. e qRT-PCR analysis of the expression of 7 candidates in the EJ and T24 T lysates after biotin-BCRC-3 pull-down assay. f RNA-FISH detection of BCRC-3 in T24 T cells. Nuclei were stained blue with DAPI. BCRC-3 was stained red with cy3. Locked nucleic acid miR-182-5p probes were labeled with Dig (scale bar, 10 μm). g qRT-PCR analysis of the expression levels of BCRC-3 or GAPDH in the EJ and T24 T lysates after biotin-miR-182-5p pull-down assay. GAPDH was used as negative control. h The luciferase activities of wild type p27 3’UTR and mutant p27 3’UTR after transfection with miR-182-5p mimic in EJ or T24 T cell lines. i j qRT-PCR and western blot analysis of the expression levels of p27 in EJ or T24 T cells after co-transfection with BCRC-3 overexpression vector and miR-182-5p mimics. (Data are mean ± SEM of three experiments. Student’s t- test analyzed the difference in a - b , d - e , g - i . * P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was carried out using the SYBR Premix Ex TaqTM kit (Takara).

    Techniques: Over Expression, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Sequencing, Pull Down Assay, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Negative Control, Quantitative RT-PCR, Expressing, Fluorescence In Situ Hybridization, Staining, Labeling, Mutagenesis, Western Blot, Cotransfection

    Alteration of HECA homo expression in HCC cell lines detected by qRT-PCR and Western blotting. (A) The HECA homo mRNA level was the highest in the HepG2 cell lines among the five examined cell lines. (B) For each of the three cell lines (HepG2, Huh-7, and MHCC-97H), the HECA homo mRNA level in the siRNA group (cells transfected with specific siRNA against HECA homo) was significantly lower than in the SNC group (cells that were transfected with scrambled siRNA as a negative control) or in the blank group (cells treated with only Lipofectamine 2000). In contrast, there was no significant difference between the SNC group and the blank group. (C) The HECA homo mRNA level in the HECA exp group (cells that were transfected with HECA homo full-length-expressing plasmid) was significantly higher than those in the mock group (cells that were transfected with empty control plasmid) or blank group (cells that were treated with only Lipofectamine 2000). Meanwhile, there was no significant difference between the mock group and the blank group. (D) and (E) the corresponding protein levels were detected by western blotting.

    Journal: PLoS ONE

    Article Title: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0137579

    Figure Lengend Snippet: Alteration of HECA homo expression in HCC cell lines detected by qRT-PCR and Western blotting. (A) The HECA homo mRNA level was the highest in the HepG2 cell lines among the five examined cell lines. (B) For each of the three cell lines (HepG2, Huh-7, and MHCC-97H), the HECA homo mRNA level in the siRNA group (cells transfected with specific siRNA against HECA homo) was significantly lower than in the SNC group (cells that were transfected with scrambled siRNA as a negative control) or in the blank group (cells treated with only Lipofectamine 2000). In contrast, there was no significant difference between the SNC group and the blank group. (C) The HECA homo mRNA level in the HECA exp group (cells that were transfected with HECA homo full-length-expressing plasmid) was significantly higher than those in the mock group (cells that were transfected with empty control plasmid) or blank group (cells that were treated with only Lipofectamine 2000). Meanwhile, there was no significant difference between the mock group and the blank group. (D) and (E) the corresponding protein levels were detected by western blotting.

    Article Snippet: Quantitative real-time PCR analysis (qRT-PCR) The total cellular RNA was extracted from cells using the MiniBest universal RNA extraction kit (Takara, Dalian, China) and reversed transcribed by the PrimeScript 1st strand cDNA synthesis kit (Takara, Dalian, China) according to the manufacturer’s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Negative Control, Plasmid Preparation

    Analysis of proline and P5CS1 . (A) qRT-PCR analysis of P5CS1 (LOC_Os01g62900). (B) Proline content under stress treatment. Data are means of three biological replicates and error bars are ± SE from three independent experiments, each performed with 6–8 leaves from five separate plants. Asterisks indicate significant differences by Tukey LSD test ( ∗ P

    Journal: Frontiers in Genetics

    Article Title: Physiological and Transcriptome Analyses Reveal Short-Term Responses and Formation of Memory Under Drought Stress in Rice

    doi: 10.3389/fgene.2019.00055

    Figure Lengend Snippet: Analysis of proline and P5CS1 . (A) qRT-PCR analysis of P5CS1 (LOC_Os01g62900). (B) Proline content under stress treatment. Data are means of three biological replicates and error bars are ± SE from three independent experiments, each performed with 6–8 leaves from five separate plants. Asterisks indicate significant differences by Tukey LSD test ( ∗ P

    Article Snippet: The relative expression levels of individual genes were measured with gene-specific primers by real-time quantitative PCR (qRT-PCR) analysis, which was carried out in a 20 μl reaction mix with 1 μl of diluted cDNA template and SYBR Premix Ex TaqII (Takara, Japan) with a Bio-Rad CFX96.

    Techniques: Quantitative RT-PCR

    MCL inhibits LPS-triggered cytokine production in Raw264.7. Plate seeding (2 × 10 5 /well) was carried out in 24-well plates overnight. Cells were stimulated by different concentrations of MCL with or without LPS (100 ng/mL) for the indicated time periods. IL-6 ( a ), TNF-α ( b ), MCP-1 ( d ), IFN-β ( e ) and IL-10 ( f ) in the supernatants were measured by ELISA. The IL-1β ( c ) mRNA expression was examined by qRT-PCR. Data are shown as mean ± SD of three independent experiments; *p

    Journal: Scientific Reports

    Article Title: Micheliolide inhibits LPS-induced inflammatory response and protects mice from LPS challenge

    doi: 10.1038/srep23240

    Figure Lengend Snippet: MCL inhibits LPS-triggered cytokine production in Raw264.7. Plate seeding (2 × 10 5 /well) was carried out in 24-well plates overnight. Cells were stimulated by different concentrations of MCL with or without LPS (100 ng/mL) for the indicated time periods. IL-6 ( a ), TNF-α ( b ), MCP-1 ( d ), IFN-β ( e ) and IL-10 ( f ) in the supernatants were measured by ELISA. The IL-1β ( c ) mRNA expression was examined by qRT-PCR. Data are shown as mean ± SD of three independent experiments; *p

    Article Snippet: Quantitative real-time RT-PCR (qRT-PCR) analysis was performed with the SYBR RT-PCR Kit (Takara, Dalian, China) and LightCycler (Roche Diagnostics, Indianapolis, IN) as described previously .

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

    Response of CdSAMDC1 transcript to cold treatment and analysis of transgenic centipedegrass (23, 24) overexpressing CdSAMDC1 in comparison to the wild type (WT). Relative expression of CdSAMDC1 in bermudagrass in response to cold (6°C) was determined by using real time quantitative RT-PCR (qRT-PCR, A ). 20 μg of genomic DNA from transgenic centipedegrass and WT plants was digested with Hind III for DNA hybridization. (B) Relative expression of CdSAMDC1 in transgenic centipedegrass was determined by using qRT-PCR using actin as reference. (C) Putrescine (Put), spermidine (Spd), and spermine (Spm) were determined by using HPLC (D) . Means of three repeats and standard errors are presented; the same letter above the column indicates no significant difference at P

    Journal: Frontiers in Plant Science

    Article Title: Transgenic Centipedegrass (Eremochloa ophiuroides [Munro] Hack.) Overexpressing S-Adenosylmethionine Decarboxylase (SAMDC) Gene for Improved Cold Tolerance Through Involvement of H2O2 and NO Signaling

    doi: 10.3389/fpls.2017.01655

    Figure Lengend Snippet: Response of CdSAMDC1 transcript to cold treatment and analysis of transgenic centipedegrass (23, 24) overexpressing CdSAMDC1 in comparison to the wild type (WT). Relative expression of CdSAMDC1 in bermudagrass in response to cold (6°C) was determined by using real time quantitative RT-PCR (qRT-PCR, A ). 20 μg of genomic DNA from transgenic centipedegrass and WT plants was digested with Hind III for DNA hybridization. (B) Relative expression of CdSAMDC1 in transgenic centipedegrass was determined by using qRT-PCR using actin as reference. (C) Putrescine (Put), spermidine (Spd), and spermine (Spm) were determined by using HPLC (D) . Means of three repeats and standard errors are presented; the same letter above the column indicates no significant difference at P

    Article Snippet: After diluted for 50-fold, the cDNA was used as template for real-time quantitative RT-PCR (qRT-PCR) analysis in a total of 10 μl PCR reaction, containing 15 ng of cDNA, 200 nM each of forward and reverse primers, and 5 μl SYBR Premix Ex Taq (Takara Bio, Inc., Otsu, Shiga, Japan), with three technical replicates and two biological replicates.

    Techniques: Transgenic Assay, Expressing, Quantitative RT-PCR, DNA Hybridization, High Performance Liquid Chromatography

    Relative mRNA levels of 9 MaAQP genes in BX and FJ were determined by qRT-PCR analysis. ( A – C ) expression patterns of MaPIP2-7, MaPIP1-4 and MaSIP1-1 in different tissues of BX and FJ. The mRNA fold difference was relative to that of BX-root samples used as calibrator; ( D – F ) expression patterns of MaPIP1-6, MaPIP2-10 and MaTIP4-1 in different stages of fruit development and ripening in BX and FJ. The mRNA fold difference was relative to that of BX-0DAF samples used as calibrator; ( G – I ) expression patterns of MaPIP2-6, MaTIP2-1 and MaTIP4-2 in response to cold, salt and osmotic stresses in BX and FJ. The mRNA fold difference was relative to that of untreated samples used as calibrator. Log 2 -based values were used to display differential expression results. Data are means ± SD of n = 3 biological replicates.

    Journal: International Journal of Molecular Sciences

    Article Title: Genome-Wide Identification and Expression Analyses of Aquaporin Gene Family during Development and Abiotic Stress in Banana

    doi: 10.3390/ijms160819728

    Figure Lengend Snippet: Relative mRNA levels of 9 MaAQP genes in BX and FJ were determined by qRT-PCR analysis. ( A – C ) expression patterns of MaPIP2-7, MaPIP1-4 and MaSIP1-1 in different tissues of BX and FJ. The mRNA fold difference was relative to that of BX-root samples used as calibrator; ( D – F ) expression patterns of MaPIP1-6, MaPIP2-10 and MaTIP4-1 in different stages of fruit development and ripening in BX and FJ. The mRNA fold difference was relative to that of BX-0DAF samples used as calibrator; ( G – I ) expression patterns of MaPIP2-6, MaTIP2-1 and MaTIP4-2 in response to cold, salt and osmotic stresses in BX and FJ. The mRNA fold difference was relative to that of untreated samples used as calibrator. Log 2 -based values were used to display differential expression results. Data are means ± SD of n = 3 biological replicates.

    Article Snippet: 4.5. qRT-PCR Analysis Changes in the expression of MaAQP genes in different tissues, different stages of fruit development and ripening, and response to abiotic stress were detected by quantitative real-time polymerase chain reaction (qRT-PCR) analysis using SYBR® Premix Ex Taq™ (TaKaRa, Shiga, Japan) chemistry on a Stratagene Mx3000P (Stratagene, CA, USA) instrument.

    Techniques: Quantitative RT-PCR, Expressing

    Expression patterns of OsCTZFP8 transcript in response to abiotic stresses. qRT-PCR was performed with 2-week-old NT plants exposed to cold (4°C), ABA (5 μ M), and NaCl (250 mM) treatments at different time points. The expression of eEF1α was used as an internal control. Data present the means ± SE of two biological replicates.

    Journal: International Journal of Genomics

    Article Title: Overexpression of a New Zinc Finger Protein Transcription Factor OsCTZFP8 Improves Cold Tolerance in Rice

    doi: 10.1155/2018/5480617

    Figure Lengend Snippet: Expression patterns of OsCTZFP8 transcript in response to abiotic stresses. qRT-PCR was performed with 2-week-old NT plants exposed to cold (4°C), ABA (5 μ M), and NaCl (250 mM) treatments at different time points. The expression of eEF1α was used as an internal control. Data present the means ± SE of two biological replicates.

    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis For qRT-PCR analysis, total RNA was isolated from abiotic stress-treated rice shoots and roots using a MiniBEST Universal RNA Extraction Kit (Takara Bio Inc., Kusatsu, Japan) according to the manufacturer's instructions.

    Techniques: Expressing, Quantitative RT-PCR

    Spatial expression pattern of OsCTZFP8 . qRT-PCR was performed with total RNA isolated from different organs of wild-type rice under normal conditions. The expression of eEF1α was used as an internal control. Data present the means ± SE of two biological replicates.

    Journal: International Journal of Genomics

    Article Title: Overexpression of a New Zinc Finger Protein Transcription Factor OsCTZFP8 Improves Cold Tolerance in Rice

    doi: 10.1155/2018/5480617

    Figure Lengend Snippet: Spatial expression pattern of OsCTZFP8 . qRT-PCR was performed with total RNA isolated from different organs of wild-type rice under normal conditions. The expression of eEF1α was used as an internal control. Data present the means ± SE of two biological replicates.

    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis For qRT-PCR analysis, total RNA was isolated from abiotic stress-treated rice shoots and roots using a MiniBEST Universal RNA Extraction Kit (Takara Bio Inc., Kusatsu, Japan) according to the manufacturer's instructions.

    Techniques: Expressing, Quantitative RT-PCR, Isolation

    Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using qRT-PCR. b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    doi: 10.1186/s13046-018-0847-7

    Figure Lengend Snippet: Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using qRT-PCR. b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed using SYBR® Premix Ex Taq™ II (Takara) on ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) in accordance with the manufacturers’ instructions.

    Techniques: Expressing, Stable Transfection, Transfection, shRNA, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Migration, Transwell Assay

    HEGBC bound to the promoter of IL11 and activated IL-11/STAT3 signaling pathway. a The distribution of HEGBC in the cytoplasmic and nuclear fractions of NOZ cells was detected using cytoplasmic and nuclear RNA isolation followed by qRT-PCR. U6 and β-actin were used as nuclear and cytoplasmic controls, respectively. b Schematic outline of the predicted binding site for HEGBC on the promoter of IL11 . c ChIRP assay in NOZ cells was performed with antisense probe sets against HEGBC or LacZ (negative control). The bound DNA was detected using qRT-PCR with specific primers against the promoters of IL11 or ACTB . d The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. e The expression of IL-11 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. f The concentration of IL11 in the culture medium from HEGBC stably overexpressed and control NOZ cells was detected using ELISA. g The concentration of IL11 in the culture medium from HEGBC stably depleted and control EH-GB2 cells was detected using ELISA. h STAT3 phosphorylation level in HEGBC stably overexpressed and control NOZ cells was detected using western blot. i STAT3 phosphorylation level in HEGBC stably depleted and control EH-GB2 cells was detected using western blot. j The expression of the target genes of STAT3 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. k The expression of the target genes of STAT3 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    doi: 10.1186/s13046-018-0847-7

    Figure Lengend Snippet: HEGBC bound to the promoter of IL11 and activated IL-11/STAT3 signaling pathway. a The distribution of HEGBC in the cytoplasmic and nuclear fractions of NOZ cells was detected using cytoplasmic and nuclear RNA isolation followed by qRT-PCR. U6 and β-actin were used as nuclear and cytoplasmic controls, respectively. b Schematic outline of the predicted binding site for HEGBC on the promoter of IL11 . c ChIRP assay in NOZ cells was performed with antisense probe sets against HEGBC or LacZ (negative control). The bound DNA was detected using qRT-PCR with specific primers against the promoters of IL11 or ACTB . d The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. e The expression of IL-11 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. f The concentration of IL11 in the culture medium from HEGBC stably overexpressed and control NOZ cells was detected using ELISA. g The concentration of IL11 in the culture medium from HEGBC stably depleted and control EH-GB2 cells was detected using ELISA. h STAT3 phosphorylation level in HEGBC stably overexpressed and control NOZ cells was detected using western blot. i STAT3 phosphorylation level in HEGBC stably depleted and control EH-GB2 cells was detected using western blot. j The expression of the target genes of STAT3 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. k The expression of the target genes of STAT3 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed using SYBR® Premix Ex Taq™ II (Takara) on ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) in accordance with the manufacturers’ instructions.

    Techniques: Isolation, Quantitative RT-PCR, Binding Assay, Negative Control, Expressing, Stable Transfection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    p-STAT3 bound to the promoter of HEGBC and activated HEGBC expression. a Schematic outline of the predicted binding site for p-STAT3 on the promoter of HEGBC . b ChIP assay in NOZ cells was performed using p-STAT3 specific antibody or negative control IgG. The bound DNA was detected using qRT-PCR with specific primers against the predicted binding sites on the promoter of HEGBC or a distal non-binding site (negative control, NC). c RIP assay in NOZ cells was performed using p-STAT3 specific antibody, STAT3 specific antibody, or negative control IgG. The bound RNA was detected using qRT-PCR with specific primers against HEGBC or TSLNC8 (positive control). d After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 5 μM p-STAT3 inhibitor SC144 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. e After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 20 ng/mL IL-11 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. f The expression of IL-11 in NOZ cells treated with 5 μM SC144 for 72 h was detected using qRT-PCR. g The expression of IL-11 in NOZ cells treated with 20 ng/mL IL-11 for 72 h was detected using qRT-PCR. For b - g , results are shown as mean ± s.d. of 3 independent experiments. ns, not significant, ** P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    doi: 10.1186/s13046-018-0847-7

    Figure Lengend Snippet: p-STAT3 bound to the promoter of HEGBC and activated HEGBC expression. a Schematic outline of the predicted binding site for p-STAT3 on the promoter of HEGBC . b ChIP assay in NOZ cells was performed using p-STAT3 specific antibody or negative control IgG. The bound DNA was detected using qRT-PCR with specific primers against the predicted binding sites on the promoter of HEGBC or a distal non-binding site (negative control, NC). c RIP assay in NOZ cells was performed using p-STAT3 specific antibody, STAT3 specific antibody, or negative control IgG. The bound RNA was detected using qRT-PCR with specific primers against HEGBC or TSLNC8 (positive control). d After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 5 μM p-STAT3 inhibitor SC144 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. e After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 20 ng/mL IL-11 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. f The expression of IL-11 in NOZ cells treated with 5 μM SC144 for 72 h was detected using qRT-PCR. g The expression of IL-11 in NOZ cells treated with 20 ng/mL IL-11 for 72 h was detected using qRT-PCR. For b - g , results are shown as mean ± s.d. of 3 independent experiments. ns, not significant, ** P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed using SYBR® Premix Ex Taq™ II (Takara) on ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) in accordance with the manufacturers’ instructions.

    Techniques: Expressing, Binding Assay, Chromatin Immunoprecipitation, Negative Control, Quantitative RT-PCR, Positive Control, Cotransfection, Luciferase, Activity Assay

    Ectopic expression of HEGBC promoted GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably overexpressed and control SGC-996 cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably overexpressed and control SGC-996 cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably overexpressed and control NOZ cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably overexpressed and control SGC-996 and NOZ cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    doi: 10.1186/s13046-018-0847-7

    Figure Lengend Snippet: Ectopic expression of HEGBC promoted GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably overexpressed and control SGC-996 cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably overexpressed and control SGC-996 cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably overexpressed and control NOZ cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably overexpressed and control SGC-996 and NOZ cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed using SYBR® Premix Ex Taq™ II (Takara) on ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) in accordance with the manufacturers’ instructions.

    Techniques: Expressing, Migration, In Vitro, Stable Transfection, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Transwell Assay

    Depletion of HEGBC inhibited GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably depleted and control GBC-SD cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably depleted and control GBC-SD cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably depleted and control EH-GB2 cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably depleted and control GBC-SD and EH-GB2 cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    doi: 10.1186/s13046-018-0847-7

    Figure Lengend Snippet: Depletion of HEGBC inhibited GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably depleted and control GBC-SD cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably depleted and control GBC-SD cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably depleted and control EH-GB2 cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably depleted and control GBC-SD and EH-GB2 cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) analyses were performed using SYBR® Premix Ex Taq™ II (Takara) on ABI StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) in accordance with the manufacturers’ instructions.

    Techniques: Migration, In Vitro, Expressing, Stable Transfection, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Transwell Assay

    qRT-PCR validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.

    Journal: International Journal of Genomics

    Article Title: Transcriptome and Expression Profiling Analysis of Recalcitrant Tea (Camellia sinensis L.) Seeds Sensitive to Dehydration

    doi: 10.1155/2018/5963797

    Figure Lengend Snippet: qRT-PCR validation of 12 selected differentially expressed genes (DEGs) responding to tea seed dehydration treatment. Validation of RNA-Seq results using qRT-PCR; GAPDH gene was chosen as the reference gene.

    Article Snippet: To validate the expression profiles observed in RNA-Seq data, 12 significantly differentially expressed genes (DEGs) were selected for qRT-PCR analyses using SYBR Premix Ex Taq™ II Kit (Takara, Dalian, China) in a Bio-Rad CFX96 real-time PCR system (Bio-Rad, CA, USA).

    Techniques: Quantitative RT-PCR, RNA Sequencing Assay

    Differences of testicular gene expression between Tex101 +/+ and Tex101 −/− mice. Analysis of Tex101 , Ly6k , Dpep3 , and Adam3 mRNA expression in the Tex101 −/− mice ( A ). Levels of these mRNA expressions in the testes of Tex101 +/+ (n = 3) and Tex101 −/− (n = 4) mice were measured using qRT-PCR with SYBR Green method. To obtain the ΔΔCt values for the calculation of fold increases, β-actin mRNA was used as a quantitative internal control. Average of each mRNA expression level in Tex101 +/+ mouse testis was defined as “relative expression value = 1.0”. The data are indicated with SE. ND: not detectable. Polysome profiles (254 nm absorbance profile) of sucrose gradient sedimentation of cell extracts from the Tex101 +/+ and Tex101 −/− mice testes ( B ). Detergent-treated cell extracts were fractionated over a 15–60% sucrose density gradient into eight fractions of equal volume. Expression levels of testicular Tex101 , Ly6k , Dpep3 , Adam3 , and β-actin mRNAs from the 8-week Tex101 +/+ and 8- or 30-week-old Tex101 −/− mice in each fraction of the sucrose gradients ( C ). Each fraction number corresponds to that in the absorbance profile shown in ( B ). Data are graphically represented as ratio of transcripts from a typical experiment, present in each fraction to the sum of all fractions. As an external standard, r-luc mRNA was used.

    Journal: Scientific Reports

    Article Title: TEX101, a glycoprotein essential for sperm fertility, is required for stable expression of Ly6k on testicular germ cells

    doi: 10.1038/srep23616

    Figure Lengend Snippet: Differences of testicular gene expression between Tex101 +/+ and Tex101 −/− mice. Analysis of Tex101 , Ly6k , Dpep3 , and Adam3 mRNA expression in the Tex101 −/− mice ( A ). Levels of these mRNA expressions in the testes of Tex101 +/+ (n = 3) and Tex101 −/− (n = 4) mice were measured using qRT-PCR with SYBR Green method. To obtain the ΔΔCt values for the calculation of fold increases, β-actin mRNA was used as a quantitative internal control. Average of each mRNA expression level in Tex101 +/+ mouse testis was defined as “relative expression value = 1.0”. The data are indicated with SE. ND: not detectable. Polysome profiles (254 nm absorbance profile) of sucrose gradient sedimentation of cell extracts from the Tex101 +/+ and Tex101 −/− mice testes ( B ). Detergent-treated cell extracts were fractionated over a 15–60% sucrose density gradient into eight fractions of equal volume. Expression levels of testicular Tex101 , Ly6k , Dpep3 , Adam3 , and β-actin mRNAs from the 8-week Tex101 +/+ and 8- or 30-week-old Tex101 −/− mice in each fraction of the sucrose gradients ( C ). Each fraction number corresponds to that in the absorbance profile shown in ( B ). Data are graphically represented as ratio of transcripts from a typical experiment, present in each fraction to the sum of all fractions. As an external standard, r-luc mRNA was used.

    Article Snippet: Analysis by qRT-PCR system To synthesize cDNA, total RNA (100–620 ng) or RNA isolated from each gradient fraction (200 ng) was reverse transcripted using the PrimeScript RT regent Kit (Takara, Shiga, Japan).

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR, SYBR Green Assay, Sedimentation

    mRNA expression analysis of six genes from the bladder TICs TF-gene regulatory network in 24 pairs of bladder cancer CD44 high vs. CD44 low tissues ETS1 , MYC , EGR1 , BAX , SLC29A6 , and ITGB1 mRNA expression was analyzed in bladder cancer CD44 high vs. CD44 low tissues using PCR (A) and qRT-PCR (B) . * p

    Journal: Oncotarget

    Article Title: Gene expression profiling and construction of a putative gene regulatory network of bladder cancer tumor-initiating cells

    doi: 10.18632/oncotarget.22771

    Figure Lengend Snippet: mRNA expression analysis of six genes from the bladder TICs TF-gene regulatory network in 24 pairs of bladder cancer CD44 high vs. CD44 low tissues ETS1 , MYC , EGR1 , BAX , SLC29A6 , and ITGB1 mRNA expression was analyzed in bladder cancer CD44 high vs. CD44 low tissues using PCR (A) and qRT-PCR (B) . * p

    Article Snippet: Real-time quantitative RT-PCR For real-time qRT-PCR analyses, less than 5 mg total RNA was reverse-transcribed into cDNA using the 1st strand cDNA synthesis kit (Takara, Dalian, China). mRNA expression for human ETS1 , EGR1 , MYC , BAX , SLC39A6 , and ITGB1 genes was examined by qRT-PCR using SYBR Premix Ex Taq (Takara) and a Biosystems 7300 Fast Real-Time PCR System.

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

    (A) Changes in AsA levels and (B, C) mRNA levels of the genes involved in AsA biosynthesis ( FaGME, FaG1PP, FaGalUR, FaGLDH , and FaMYOX ) and recycling ( FaMDHAR and FaDHAR ) by QRT-PCR. (D) FaGalUR protein levels in the ripe fruits of the parentals (1392 and 232) as well as lines with high (93-30 and 93-34), medium (93-29), and low (93-67 and 93-72) AsA levels in the segregating population. The QRT-PCR values were normalized against the abundance of FaRib413 , using the parental 1392 value as the reference (value 1). The bars represent the mean of at least two independent biological samples ±SD. Different letters indicate a significant difference between samples according to the corresponding ANOVA ( P

    Journal: Journal of Experimental Botany

    Article Title: Regulation of L-ascorbic acid content in strawberry fruits

    doi: 10.1093/jxb/err122

    Figure Lengend Snippet: (A) Changes in AsA levels and (B, C) mRNA levels of the genes involved in AsA biosynthesis ( FaGME, FaG1PP, FaGalUR, FaGLDH , and FaMYOX ) and recycling ( FaMDHAR and FaDHAR ) by QRT-PCR. (D) FaGalUR protein levels in the ripe fruits of the parentals (1392 and 232) as well as lines with high (93-30 and 93-34), medium (93-29), and low (93-67 and 93-72) AsA levels in the segregating population. The QRT-PCR values were normalized against the abundance of FaRib413 , using the parental 1392 value as the reference (value 1). The bars represent the mean of at least two independent biological samples ±SD. Different letters indicate a significant difference between samples according to the corresponding ANOVA ( P

    Article Snippet: Expression was analysed by quantitative RT-PCR (qRT-PCR) using a SYBR® Premix Ex Taq™ sample (Takara, http://www.takara-bio.com/ ), a Rotor Gene detection system (Qiagen, http://www.qiagen.com ) according to the manufacturer's instructions, and gene-specific primers (see Supplementary Table S1 at JXB online).

    Techniques: Quantitative RT-PCR

    Comparative qRT-PCR analysis of mRNA levels of the Kctd14 gene between Znf230 KO and C57BL/6J (WT) mice. Bars represent the means ± S.D., Statistical p-values

    Journal: Genetics and Molecular Biology

    Article Title: Targeted disruption of the mouse testis-enriched gene Znf230 does not affect spermatogenesis or fertility

    doi: 10.1590/S1415-47572014005000013

    Figure Lengend Snippet: Comparative qRT-PCR analysis of mRNA levels of the Kctd14 gene between Znf230 KO and C57BL/6J (WT) mice. Bars represent the means ± S.D., Statistical p-values

    Article Snippet: Quantitative real time PCR (qRT-PCR) qRT-PCR analyses were performed with cDNA using a SYBR® Premix Ex TaqTM II kit (Takara, Dalan, China) and 10 mM of the corresponding set of sense and antisense primers of the Kctd14 gene: 5′-atgggcaccctgatgaagc -3′ (Forward) and 5′-gcccagtgcgcaggtagtc -3′ (Reverse).

    Techniques: Quantitative RT-PCR, Mouse Assay

    ABI3 regulates the expression of TIP3 genes. (A and B) qRT-PCR and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.

    Journal: Journal of Experimental Botany

    Article Title: Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3

    doi: 10.1093/jxb/erv244

    Figure Lengend Snippet: ABI3 regulates the expression of TIP3 genes. (A and B) qRT-PCR and immunoblot analysis of the expression of TIP3 genes in abi3-6 and fus3-3 seeds. Values in (A) are means ±SD, n =3. (C) The TIP3;1 and TIP3;2 promoters are activated by ABI3 when treated with ABA in a transient expression assay. Values are means ±SD, n =3. Protoplasts transformed with empty pGREENII 62-SK vector were used as a control; 5 μM ABA was supplied in the ABA treatment. (D) qRT-PCR analysis of TIP3 and EM1 transcript levels in the WT (Col) and a transgenic line ectopically expressing ABI3 ( Pro 35S :ABI3 ). For ABA treatment, 3-week-old seedlings grown on MS medium were transferred to MS medium supplemented with 50 μM ABA for 3 d. PP2A was used as an endogenous control. (E) Immunoblot analysis of TIP3s and TIP1s in protoplasts transiently expressing ABI3 or FUS3 in the presence of 5 μM ABA. Detection of actin by an antibody was used as a loading control. (F) Immunoblot analysis of TIP3s in seedlings of WT and Pro 35S :ABI3 transgenic Arabidopsis . ABA treatment was performed as described in (D). DS, dry mature seeds.

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analyses were performed using the SYBR Green method (SYBR premix EX taq, TaKaRa) with the StepOnePlus™ Real-time PCR System (Applied Biosystems).

    Techniques: Expressing, Quantitative RT-PCR, Transformation Assay, Plasmid Preparation, Transgenic Assay, Mass Spectrometry

    Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1 -RNAi transgenic lines ( TIP3;1 -RNAi/ tip3;2 ) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1 , tip3;2 , and tip3;1/tip3;2 . LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1 , TIP3;2 , and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1 , TIP3;2 , and ACT7 was quantified by comparisons with that of PP2A . Values are means ±SD, n =3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1 -RNAi/ tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)

    Journal: Journal of Experimental Botany

    Article Title: Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3

    doi: 10.1093/jxb/erv244

    Figure Lengend Snippet: Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1 -RNAi transgenic lines ( TIP3;1 -RNAi/ tip3;2 ) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1 , tip3;2 , and tip3;1/tip3;2 . LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1 , TIP3;2 , and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1 , TIP3;2 , and ACT7 was quantified by comparisons with that of PP2A . Values are means ±SD, n =3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1 -RNAi/ tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analyses were performed using the SYBR Green method (SYBR premix EX taq, TaKaRa) with the StepOnePlus™ Real-time PCR System (Applied Biosystems).

    Techniques: Transgenic Assay, Mutagenesis, Genomic Sequencing, Polymerase Chain Reaction, Construct, Quantitative RT-PCR

    TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis . qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n =3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.

    Journal: Journal of Experimental Botany

    Article Title: Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3

    doi: 10.1093/jxb/erv244

    Figure Lengend Snippet: TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis . qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n =3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.

    Article Snippet: Quantitative RT-PCR (qRT-PCR) analyses were performed using the SYBR Green method (SYBR premix EX taq, TaKaRa) with the StepOnePlus™ Real-time PCR System (Applied Biosystems).

    Techniques: Expressing, Quantitative RT-PCR, SDS Page, Staining