quantitative pcr Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • N/A
    The One Step RT PCR Kit provides an efficient accurate and convenient way to synthesize cDNA and amplify via PCR from any template compatible with both Poly A mRNA and
      Buy from Supplier

    N/A
    Verified forward and reverse primers for analyzing the quantitative expression of gene
      Buy from Supplier

    99
    Thermo Fisher quantitative real time pcr
    Quantitative Real Time Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 45481 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr/product/Thermo Fisher
    Average 99 stars, based on 45481 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Bio-Rad qpcr
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Qpcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 11296 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qpcr/product/Bio-Rad
    Average 99 stars, based on 11296 article reviews
    Price from $9.99 to $1999.99
    qpcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    94
    Promega gotaq qpcr master mix
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Gotaq Qpcr Master Mix, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 11162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gotaq qpcr master mix/product/Promega
    Average 94 stars, based on 11162 article reviews
    Price from $9.99 to $1999.99
    gotaq qpcr master mix - by Bioz Stars, 2020-12
    94/100 stars
      Buy from Supplier

    99
    Bio-Rad rt qpcr
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Rt Qpcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 5607 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rt qpcr/product/Bio-Rad
    Average 99 stars, based on 5607 article reviews
    Price from $9.99 to $1999.99
    rt qpcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher qpcr
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22021 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qpcr/product/Thermo Fisher
    Average 99 stars, based on 22021 article reviews
    Price from $9.99 to $1999.99
    qpcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    93
    Toyobo thunderbird sybr qpcr mix
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Thunderbird Sybr Qpcr Mix, supplied by Toyobo, used in various techniques. Bioz Stars score: 93/100, based on 8167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/thunderbird sybr qpcr mix/product/Toyobo
    Average 93 stars, based on 8167 article reviews
    Price from $9.99 to $1999.99
    thunderbird sybr qpcr mix - by Bioz Stars, 2020-12
    93/100 stars
      Buy from Supplier

    95
    Toyobo revertra ace qpcr rt kit
    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative <t>PCR</t> and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P
    Revertra Ace Qpcr Rt Kit, supplied by Toyobo, used in various techniques. Bioz Stars score: 95/100, based on 8890 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/revertra ace qpcr rt kit/product/Toyobo
    Average 95 stars, based on 8890 article reviews
    Price from $9.99 to $1999.99
    revertra ace qpcr rt kit - by Bioz Stars, 2020-12
    95/100 stars
      Buy from Supplier

    96
    Bio-Rad quantitative real time pcr
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Quantitative Real Time Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 15629 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr/product/Bio-Rad
    Average 96 stars, based on 15629 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr - by Bioz Stars, 2020-12
    96/100 stars
      Buy from Supplier

    99
    Thermo Fisher platinum sybr green qpcr supermix udg
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Platinum Sybr Green Qpcr Supermix Udg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10276 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum sybr green qpcr supermix udg/product/Thermo Fisher
    Average 99 stars, based on 10276 article reviews
    Price from $9.99 to $1999.99
    platinum sybr green qpcr supermix udg - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher maxima first strand cdna synthesis kit
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Maxima First Strand Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9764 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maxima first strand cdna synthesis kit/product/Thermo Fisher
    Average 99 stars, based on 9764 article reviews
    Price from $9.99 to $1999.99
    maxima first strand cdna synthesis kit - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher real time quantitative pcr
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Real Time Quantitative Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 21195 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time quantitative pcr/product/Thermo Fisher
    Average 99 stars, based on 21195 article reviews
    Price from $9.99 to $1999.99
    real time quantitative pcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher maxima sybr green rox qpcr master mix
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Maxima Sybr Green Rox Qpcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6983 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maxima sybr green rox qpcr master mix/product/Thermo Fisher
    Average 99 stars, based on 6983 article reviews
    Price from $9.99 to $1999.99
    maxima sybr green rox qpcr master mix - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    94
    Toyobo revertra ace qpcr rt master mix
    <t>IL-17A</t> does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time <t>PCR</t> analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.
    Revertra Ace Qpcr Rt Master Mix, supplied by Toyobo, used in various techniques. Bioz Stars score: 94/100, based on 5821 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/revertra ace qpcr rt master mix/product/Toyobo
    Average 94 stars, based on 5821 article reviews
    Price from $9.99 to $1999.99
    revertra ace qpcr rt master mix - by Bioz Stars, 2020-12
    94/100 stars
      Buy from Supplier

    91
    Bio-Rad quantitative reverse transcription polymerase chain reaction
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Quantitative Reverse Transcription Polymerase Chain Reaction, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative reverse transcription polymerase chain reaction/product/Bio-Rad
    Average 91 stars, based on 42 article reviews
    Price from $9.99 to $1999.99
    quantitative reverse transcription polymerase chain reaction - by Bioz Stars, 2020-12
    91/100 stars
      Buy from Supplier

    99
    Thermo Fisher maxima sybr green qpcr master mix
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Maxima Sybr Green Qpcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6413 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maxima sybr green qpcr master mix/product/Thermo Fisher
    Average 99 stars, based on 6413 article reviews
    Price from $9.99 to $1999.99
    maxima sybr green qpcr master mix - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher quantitative real time pcr qpcr
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Quantitative Real Time Pcr Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4920 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr qpcr/product/Thermo Fisher
    Average 99 stars, based on 4920 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr qpcr - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    94
    Agilent technologies mx3000p qpcr system
    (A) SYBR green dissociation curve analyses of <t>LSG-qPCR</t> amplicons from the Stratagene <t>Mx3000P</t> platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis
    Mx3000p Qpcr System, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 3788 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mx3000p qpcr system/product/Agilent technologies
    Average 94 stars, based on 3788 article reviews
    Price from $9.99 to $1999.99
    mx3000p qpcr system - by Bioz Stars, 2020-12
    94/100 stars
      Buy from Supplier

    92
    Stratagene mx3000p qpcr system
    (A) SYBR green dissociation curve analyses of <t>LSG-qPCR</t> amplicons from the Stratagene <t>Mx3000P</t> platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis
    Mx3000p Qpcr System, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 4131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mx3000p qpcr system/product/Stratagene
    Average 92 stars, based on 4131 article reviews
    Price from $9.99 to $1999.99
    mx3000p qpcr system - by Bioz Stars, 2020-12
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher platinum qpcr supermix udg
    (A) SYBR green dissociation curve analyses of <t>LSG-qPCR</t> amplicons from the Stratagene <t>Mx3000P</t> platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis
    Platinum Qpcr Supermix Udg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 380 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum qpcr supermix udg/product/Thermo Fisher
    Average 99 stars, based on 380 article reviews
    Price from $9.99 to $1999.99
    platinum qpcr supermix udg - by Bioz Stars, 2020-12
    99/100 stars
      Buy from Supplier

    97
    Millipore quantitative pcr
    (A) SYBR green dissociation curve analyses of <t>LSG-qPCR</t> amplicons from the Stratagene <t>Mx3000P</t> platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis
    Quantitative Pcr, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 441 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative pcr/product/Millipore
    Average 97 stars, based on 441 article reviews
    Price from $9.99 to $1999.99
    quantitative pcr - by Bioz Stars, 2020-12
    97/100 stars
      Buy from Supplier

    N/A
    Verified forward and reverse primers for analyzing the quantitative expression of gene
      Buy from Supplier

    N/A
    Verified forward and reverse primers for analyzing the quantitative expression of gene
      Buy from Supplier

    Image Search Results


    Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative PCR and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P

    Journal: Genome Biology

    Article Title: Defective structural RNA processing in relapsing-remitting multiple sclerosis

    doi: 10.1186/s13059-015-0629-x

    Figure Lengend Snippet: Extensive mRNA isoform loss and intron retention in RRMS. ( A ) Total exon loss and intron gain events across the genome in RRMS (N = 6) and CTRL (N = 8) were determined from RNA-seq analysis (see Methods ). Results are expressed as average normalized counts or reads across known exons and introns in CTRL and RRMS. ( B ) Percent abundance of transcript alterations in RRMS relative to CTRL PBMC determined from analysis of genome-wide RNA-seq data. ( C ) Example of intron retention in mRNA encoded by MBP . Green arrows identify exons. Red circles identify retention of intron sequences in mature mRNA in RRMS. Orange and blue arrows identify two isoforms. ( D , E ) Examples of 5’ mRNA shortening, CSF1R , and 3’ mRNA shortening, NFATC1 in RRMS. Red arrows identify loss of exons in RRMS and green arrows identify exons expressed at equal levels between CTRL and RRMS. ( F ) Expression levels of individual exons and the MBP intron in CTRL (N = 12) and RRMS (N = 12) was determined by quantitative PCR and normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. * = P

    Article Snippet: Real-time qPCR (Bio-Rad iCycleriQ Real Time PCR System) was performed in duplicate using SYBR green in 15 μL reaction volumes.

    Techniques: RNA Sequencing Assay, Genome Wide, Expressing, Real-time Polymerase Chain Reaction

    Loss of TROVE2 and/or SSB via small RNA interference increases poly(A) + Y1 RNA, rRNAs, and U1 RNA, misprocessed 18S and 28S rRNA, and alters mRNA length. ( A ) Selective siRNA-mediated knockdown of TROVE2 or SSB in THP-1 cells or Jurkat T cells. Transcript levels of TROVE2 and SSB were determined by quantitative PCR using oligo d(T) for cDNA synthesis. Results are normalized to cells transfected with a non-specific scrambled siRNA negative control after normalization to transcript levels of GAPDH . ( B ) Poly(A) + Y1 RNA levels were determined by quantitative PCR using oligo-dT for cDNA synthesis. Results normalized to CTRL (transfection of scrambled siRNA). ( C ) As in ( B ) except levels of 18S and 28S rRNAs were determined. ( D ) As in (B) except levels of poly(A) + U1 snRNA were determined. ( E , F ) Transcript levels of misprocessed rRNAs in THP-1 (E) and Jurkat T cells (F) (-90, -60, -30 bp) were determined and are expressed relative to cells transfected with a scrambled siRNA as described in Figure 2 . ( G ) PCR strategy to test altered lengths of MBP , CSF1R , and NFATC1 transcripts. ( H ) As in (A) except transcript levels of the MBP intron, CSF1R exons, and NFATC1 short to long mRNA isoform ratios were determined by quantitative PCR and normalized to CTRL. Each experiment was performed a minimum of three times, results are expressed as mean ± S.D * P

    Journal: Genome Biology

    Article Title: Defective structural RNA processing in relapsing-remitting multiple sclerosis

    doi: 10.1186/s13059-015-0629-x

    Figure Lengend Snippet: Loss of TROVE2 and/or SSB via small RNA interference increases poly(A) + Y1 RNA, rRNAs, and U1 RNA, misprocessed 18S and 28S rRNA, and alters mRNA length. ( A ) Selective siRNA-mediated knockdown of TROVE2 or SSB in THP-1 cells or Jurkat T cells. Transcript levels of TROVE2 and SSB were determined by quantitative PCR using oligo d(T) for cDNA synthesis. Results are normalized to cells transfected with a non-specific scrambled siRNA negative control after normalization to transcript levels of GAPDH . ( B ) Poly(A) + Y1 RNA levels were determined by quantitative PCR using oligo-dT for cDNA synthesis. Results normalized to CTRL (transfection of scrambled siRNA). ( C ) As in ( B ) except levels of 18S and 28S rRNAs were determined. ( D ) As in (B) except levels of poly(A) + U1 snRNA were determined. ( E , F ) Transcript levels of misprocessed rRNAs in THP-1 (E) and Jurkat T cells (F) (-90, -60, -30 bp) were determined and are expressed relative to cells transfected with a scrambled siRNA as described in Figure 2 . ( G ) PCR strategy to test altered lengths of MBP , CSF1R , and NFATC1 transcripts. ( H ) As in (A) except transcript levels of the MBP intron, CSF1R exons, and NFATC1 short to long mRNA isoform ratios were determined by quantitative PCR and normalized to CTRL. Each experiment was performed a minimum of three times, results are expressed as mean ± S.D * P

    Article Snippet: Real-time qPCR (Bio-Rad iCycleriQ Real Time PCR System) was performed in duplicate using SYBR green in 15 μL reaction volumes.

    Techniques: Real-time Polymerase Chain Reaction, Transfection, Negative Control, Polymerase Chain Reaction

    Increased total and polyadenylated U snRNAs in RRMS. ( A ) Total U1 RNA transcript levels were determined by quantitative PCR using random hexamers for cDNA synthesis in CTRL (N = 24) and RRMS (N = 22). Individual transcripts were normalized to GAPDH transcript levels. ( B ) As in (A) except total poly(A) + U1 snRNA levels were determined by quantitative PCR using oligo-dT for cDNA synthesis in CTRL (N = 24), CIS-MS (N = 16), RRMS (N = 22), SLE (N = 24), RA (N = 18), and NMO (N = 22). ( C ) As in (A), except transcript levels were determined by whole genome RNA-sequencing (RNA-seq) and normalized to CTRL = 1; CTRL (N = 8) and RRMS (N = 6). ( D ) As in (A, B) except transcript levels of U11 and U12 snRNAs were determined by quantitative PCR using oligo d(T) for poly(A) + or random hexamer primers for total U11 or U12 snRNA.

    Journal: Genome Biology

    Article Title: Defective structural RNA processing in relapsing-remitting multiple sclerosis

    doi: 10.1186/s13059-015-0629-x

    Figure Lengend Snippet: Increased total and polyadenylated U snRNAs in RRMS. ( A ) Total U1 RNA transcript levels were determined by quantitative PCR using random hexamers for cDNA synthesis in CTRL (N = 24) and RRMS (N = 22). Individual transcripts were normalized to GAPDH transcript levels. ( B ) As in (A) except total poly(A) + U1 snRNA levels were determined by quantitative PCR using oligo-dT for cDNA synthesis in CTRL (N = 24), CIS-MS (N = 16), RRMS (N = 22), SLE (N = 24), RA (N = 18), and NMO (N = 22). ( C ) As in (A), except transcript levels were determined by whole genome RNA-sequencing (RNA-seq) and normalized to CTRL = 1; CTRL (N = 8) and RRMS (N = 6). ( D ) As in (A, B) except transcript levels of U11 and U12 snRNAs were determined by quantitative PCR using oligo d(T) for poly(A) + or random hexamer primers for total U11 or U12 snRNA.

    Article Snippet: Real-time qPCR (Bio-Rad iCycleriQ Real Time PCR System) was performed in duplicate using SYBR green in 15 μL reaction volumes.

    Techniques: Real-time Polymerase Chain Reaction, Mass Spectrometry, RNA Sequencing Assay, Random Hexamer Labeling

    Ro60 and La proteins are depressed in RRMS. ( A ) Ro60 ( TROVE2 ) and La ( SSB ) transcript levels in CTRL (N = 24), CIS-MS (N = 16), RRMS (N = 22), RA (N = 18), SLE (N = 24), NMO (N = 22), and PD (N = 19) were determined by quantitative PCR after cDNA synthesis using oligo-dT. Results are normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. ( B ) As in (A) using whole genome RNA-sequencing data. ( C ) Western blotting to determine Ro60 and La protein levels in PBMC from CTRL (N = 9) and RRMS (N = 8). ( D ) Quantitative estimates of protein abundance relative to β-actin. * P

    Journal: Genome Biology

    Article Title: Defective structural RNA processing in relapsing-remitting multiple sclerosis

    doi: 10.1186/s13059-015-0629-x

    Figure Lengend Snippet: Ro60 and La proteins are depressed in RRMS. ( A ) Ro60 ( TROVE2 ) and La ( SSB ) transcript levels in CTRL (N = 24), CIS-MS (N = 16), RRMS (N = 22), RA (N = 18), SLE (N = 24), NMO (N = 22), and PD (N = 19) were determined by quantitative PCR after cDNA synthesis using oligo-dT. Results are normalized to CTRL = 1.0 after normalization to transcript levels of GAPDH , error bars are S.D. ( B ) As in (A) using whole genome RNA-sequencing data. ( C ) Western blotting to determine Ro60 and La protein levels in PBMC from CTRL (N = 9) and RRMS (N = 8). ( D ) Quantitative estimates of protein abundance relative to β-actin. * P

    Article Snippet: Real-time qPCR (Bio-Rad iCycleriQ Real Time PCR System) was performed in duplicate using SYBR green in 15 μL reaction volumes.

    Techniques: Mass Spectrometry, Real-time Polymerase Chain Reaction, RNA Sequencing Assay, Western Blot

    Interferon-β1b (IFN-β1b) therapy corrects aberrant levels of TROVE2 , SSB , poly (A) + Y1 RNA, and poly(A) + U1 snRNA in RRMS. Blood samples were collected in PaxGene tubes from CTRL (N = 12), RRMS subjects not on IFN-β1b (RRMS - IFN-β1b, N = 12), and RRMS subjects on stable IFN-β1b therapy (RRMS + IFN-β1b, N = 4). Oligo dT was used for cDNA synthesis. Transcript levels of TROVE2 and SSB ( A ) or poly(A) + Y1 RNA and poly(A) + U1 RNA ( B ) were determined by quantitative PCR and normalized to CTRL = 1 after normalization to levels of GAPDH . Error bars are ± S.D. * P

    Journal: Genome Biology

    Article Title: Defective structural RNA processing in relapsing-remitting multiple sclerosis

    doi: 10.1186/s13059-015-0629-x

    Figure Lengend Snippet: Interferon-β1b (IFN-β1b) therapy corrects aberrant levels of TROVE2 , SSB , poly (A) + Y1 RNA, and poly(A) + U1 snRNA in RRMS. Blood samples were collected in PaxGene tubes from CTRL (N = 12), RRMS subjects not on IFN-β1b (RRMS - IFN-β1b, N = 12), and RRMS subjects on stable IFN-β1b therapy (RRMS + IFN-β1b, N = 4). Oligo dT was used for cDNA synthesis. Transcript levels of TROVE2 and SSB ( A ) or poly(A) + Y1 RNA and poly(A) + U1 RNA ( B ) were determined by quantitative PCR and normalized to CTRL = 1 after normalization to levels of GAPDH . Error bars are ± S.D. * P

    Article Snippet: Real-time qPCR (Bio-Rad iCycleriQ Real Time PCR System) was performed in duplicate using SYBR green in 15 μL reaction volumes.

    Techniques: Real-time Polymerase Chain Reaction

    Restriction of HBV by SAMHD1 is dependent upon dNTPase activity, is inhibited by phosphorylation of T592 and is rescued by addition of deoxynucleosides. ( a , b ) HepG2.2.15 cells were transfected with 1 μg of plasmids coding for wild-type or mutant FLAG-tagged SAMHD1 or with 1 μg of empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements for 48 hours. ( a ) Detection of overexpressed FLAG-SAMHD1 by Western blotting. Detection of actin was used as a loading control. ( b ) HBV DNA from the supernatant was quantified by qPCR. The mean ± SEM of technical triplicates from one representative experiment out of three independent experiments is represented. ( c ) HepG2 cells were transfected with 1 μg of plasmid coding for wild-type FLAG-tagged SAMHD1 or empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements to arrest the cell cycle. When indicated, 10 mM of hydroxyurea (HU) were added at the time of medium change. Quantification of dATP levels 3 days post medium change were determined by single-nucleotide incorporation assay. The mean ± SEM of technical duplicates is represented. ( d ) HepG2.2.15 cells were transfected with 1 μg of plasmid coding for wild-type FLAG-tagged SAMHD1 or with an empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements. When indicated, deoxynucleosides (dNs, 2 mM each) or solvent were added at the time of medium change. HBV DNA from the supernatant was quantified by qPCR 3 days post medium change. For each independent experiment, fold-changes to empty vector were calculated based on the median of three technical replicates. The mean ± SEM of the fold-changes of three independent experiments is represented. ( b , d ) Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (**p

    Journal: Scientific Reports

    Article Title: Restrictive influence of SAMHD1 on Hepatitis B Virus life cycle

    doi: 10.1038/srep26616

    Figure Lengend Snippet: Restriction of HBV by SAMHD1 is dependent upon dNTPase activity, is inhibited by phosphorylation of T592 and is rescued by addition of deoxynucleosides. ( a , b ) HepG2.2.15 cells were transfected with 1 μg of plasmids coding for wild-type or mutant FLAG-tagged SAMHD1 or with 1 μg of empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements for 48 hours. ( a ) Detection of overexpressed FLAG-SAMHD1 by Western blotting. Detection of actin was used as a loading control. ( b ) HBV DNA from the supernatant was quantified by qPCR. The mean ± SEM of technical triplicates from one representative experiment out of three independent experiments is represented. ( c ) HepG2 cells were transfected with 1 μg of plasmid coding for wild-type FLAG-tagged SAMHD1 or empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements to arrest the cell cycle. When indicated, 10 mM of hydroxyurea (HU) were added at the time of medium change. Quantification of dATP levels 3 days post medium change were determined by single-nucleotide incorporation assay. The mean ± SEM of technical duplicates is represented. ( d ) HepG2.2.15 cells were transfected with 1 μg of plasmid coding for wild-type FLAG-tagged SAMHD1 or with an empty vector. Twenty-four hours after transfection, the cells were washed and cultured in serum-free medium without supplements. When indicated, deoxynucleosides (dNs, 2 mM each) or solvent were added at the time of medium change. HBV DNA from the supernatant was quantified by qPCR 3 days post medium change. For each independent experiment, fold-changes to empty vector were calculated based on the median of three technical replicates. The mean ± SEM of the fold-changes of three independent experiments is represented. ( b , d ) Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (**p

    Article Snippet: For HBV total DNA, qPCR reactions were performed on undigested samples in SYBR green supermix (Bio-rad 172-5121) using HBV specific primers HBV-total-DNA-F and HBV-total-DNA-R (95 °C for 10 minutes, followed up by 95 °C for 10 seconds and 60 °C for 30 seconds for 40 cycles).

    Techniques: Activity Assay, Transfection, Mutagenesis, Plasmid Preparation, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction

    Overexpression of SAMHD1 reduces extracellular and intracellular levels of HBV DNA in HepG2.2.15 cells without affecting viral RNA. HepG2.2.15 cells were transfected with ( a ) increasing amounts or ( b – d ) 1 μg of a SAMHD1-coding plasmid. The total amount of plasmid DNA was adjusted to 1 μg by addition of empty vector. Twenty-four hours after transfection, the cells were washed with phosphate-buffered saline (PBS) and further cultured in serum-free medium without supplements. Forty-eight hours later, HBV DNA from the supernatant ( a , d ) and intracellular HBV DNA from cytoplasmic fractions ( c ) were quantified by qPCR. ( b ) The levels of intracellular HBV RNAs were determined by RT-qPCR using primer sets that amplify the indicated HBV RNAs (3.5 kb; 3.5 kb and 2.4 kb; 3.5 kb, 2.4 kb and 2.1 kb, or all four HBV RNAs). ( a ) The mean ± SEM of the technical triplicates is shown. Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Journal: Scientific Reports

    Article Title: Restrictive influence of SAMHD1 on Hepatitis B Virus life cycle

    doi: 10.1038/srep26616

    Figure Lengend Snippet: Overexpression of SAMHD1 reduces extracellular and intracellular levels of HBV DNA in HepG2.2.15 cells without affecting viral RNA. HepG2.2.15 cells were transfected with ( a ) increasing amounts or ( b – d ) 1 μg of a SAMHD1-coding plasmid. The total amount of plasmid DNA was adjusted to 1 μg by addition of empty vector. Twenty-four hours after transfection, the cells were washed with phosphate-buffered saline (PBS) and further cultured in serum-free medium without supplements. Forty-eight hours later, HBV DNA from the supernatant ( a , d ) and intracellular HBV DNA from cytoplasmic fractions ( c ) were quantified by qPCR. ( b ) The levels of intracellular HBV RNAs were determined by RT-qPCR using primer sets that amplify the indicated HBV RNAs (3.5 kb; 3.5 kb and 2.4 kb; 3.5 kb, 2.4 kb and 2.1 kb, or all four HBV RNAs). ( a ) The mean ± SEM of the technical triplicates is shown. Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Article Snippet: For HBV total DNA, qPCR reactions were performed on undigested samples in SYBR green supermix (Bio-rad 172-5121) using HBV specific primers HBV-total-DNA-F and HBV-total-DNA-R (95 °C for 10 minutes, followed up by 95 °C for 10 seconds and 60 °C for 30 seconds for 40 cycles).

    Techniques: Over Expression, Transfection, Plasmid Preparation, Cell Culture, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    dATP levels are affected by HBV infection. SAMHD1 is de-phosphorylated at T592 in primary human hepatocytes, and is not affected by HBV infection. ( a ) HepG2-NTCP- control-shRNA cells were infected or not with HBV in presence of 2.5%DMSO and dATP content was measured 4 days post infection. The mean ± SEM of technical duplicates is represented. ( b , c ) Primary human hepatocytes (PHH) from one donnor were infected or not with HBV for 8 or 24 hours. ( b ) SAMHD1 phosphorylation on T592, total SAMHD1, cyclin B1 and GAPDH were detected by Western blotting. Cycling or serum starved HepG2.2.15-control-shRNA cells were used respectively as positive and negative controls for pT592-SAMHD1 and cyclin B1. A representative experiment out of two technical replicates is represented. ( c ) Levels of intracellular HBV RNAs from infected PHH used in ( b ) were determined by RT-qPCR using HBV primer set 3 (see Fig. 2b and Table 1 ; amplified HBV RNA species: 3.5 kb, 2.4 kb and 2.1 kb RNAs). B.d.: below detection. The mean ± SEM of technical triplicates is represented.

    Journal: Scientific Reports

    Article Title: Restrictive influence of SAMHD1 on Hepatitis B Virus life cycle

    doi: 10.1038/srep26616

    Figure Lengend Snippet: dATP levels are affected by HBV infection. SAMHD1 is de-phosphorylated at T592 in primary human hepatocytes, and is not affected by HBV infection. ( a ) HepG2-NTCP- control-shRNA cells were infected or not with HBV in presence of 2.5%DMSO and dATP content was measured 4 days post infection. The mean ± SEM of technical duplicates is represented. ( b , c ) Primary human hepatocytes (PHH) from one donnor were infected or not with HBV for 8 or 24 hours. ( b ) SAMHD1 phosphorylation on T592, total SAMHD1, cyclin B1 and GAPDH were detected by Western blotting. Cycling or serum starved HepG2.2.15-control-shRNA cells were used respectively as positive and negative controls for pT592-SAMHD1 and cyclin B1. A representative experiment out of two technical replicates is represented. ( c ) Levels of intracellular HBV RNAs from infected PHH used in ( b ) were determined by RT-qPCR using HBV primer set 3 (see Fig. 2b and Table 1 ; amplified HBV RNA species: 3.5 kb, 2.4 kb and 2.1 kb RNAs). B.d.: below detection. The mean ± SEM of technical triplicates is represented.

    Article Snippet: For HBV total DNA, qPCR reactions were performed on undigested samples in SYBR green supermix (Bio-rad 172-5121) using HBV specific primers HBV-total-DNA-F and HBV-total-DNA-R (95 °C for 10 minutes, followed up by 95 °C for 10 seconds and 60 °C for 30 seconds for 40 cycles).

    Techniques: Infection, shRNA, Western Blot, Quantitative RT-PCR, Amplification

    SAMHD1 restricts HBV replication in infected HepG2-NTCP cells. Detection of ( a ) SAMHD1 or ( b ) NTCP in SAMHD1 knockdown or control HepG2-NTCP cells by Western blotting. Actin or GAPDH were used as loading controls. ( c ) Equal cell growth and viability was assessed was assessed using ATPLite. Depicted are mean values (±SEM) of three biological replicates of one representative experiment out of three. ( d ) Control shRNA cells were cultured for 10 days in complete medium supplemented (+) or not (−) with 2.5% DMSO. When indicated cells were infected with HBV 48 hours after start of DMSO treatment. Phosphorylation at residue T592 in SAMHD1, as well as expression of total SAMHD1, cyclin B1 and actin, were detected by Western blotting. ( e ) Cells were cultured for 10 days in 2.5% DMSO-containing medium and infected with a single-round luciferase HIV-1 virus. Luciferase activity was detected 24 hours post infection. Depicted are mean values (±SEM) of three biological replicates of one representative experiment out of three. ( f-i ) Cells were infected with HBV inoculum, and where indicated, the entry inhibitor MyrcludexB (MyrB, 200 nM), or the reverse transcription inhibitor lamivudine (Lami, 0,5 μM) were added. Cells were cultured in medium containing 2.5% DMSO. Ten days post infection, HBV DNA from the supernatant ( f ), intracellular total HBV DNA ( g ) and cccDNA ( h ) were quantified by qPCR. ( i ) The intracellular HBV RNAs levels were determined 10 days post infection by RT-qPCR using primer sets that amplify the indicated HBV RNAs (3.5 kb; 3.5 kb and 2.4 kb; 3.5 kb, 2.4 kb and 2.1 kb, or all four HBV RNAs). ( f ) For each individual experiment, fold-changes to the untreated control shRNA were calculated based on the median of three biological replicates, each measured in three technical replicates. The mean ± SEM of the fold-changes of three independent experiments is depicted. ( g – i ) For each independent experiment, fold-changes to untreated control shRNA were calculated based on the median of three technical replicates. The mean ± SEM of the fold-changes of at least 3 independent experiments is represented. ( f – i ) Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Journal: Scientific Reports

    Article Title: Restrictive influence of SAMHD1 on Hepatitis B Virus life cycle

    doi: 10.1038/srep26616

    Figure Lengend Snippet: SAMHD1 restricts HBV replication in infected HepG2-NTCP cells. Detection of ( a ) SAMHD1 or ( b ) NTCP in SAMHD1 knockdown or control HepG2-NTCP cells by Western blotting. Actin or GAPDH were used as loading controls. ( c ) Equal cell growth and viability was assessed was assessed using ATPLite. Depicted are mean values (±SEM) of three biological replicates of one representative experiment out of three. ( d ) Control shRNA cells were cultured for 10 days in complete medium supplemented (+) or not (−) with 2.5% DMSO. When indicated cells were infected with HBV 48 hours after start of DMSO treatment. Phosphorylation at residue T592 in SAMHD1, as well as expression of total SAMHD1, cyclin B1 and actin, were detected by Western blotting. ( e ) Cells were cultured for 10 days in 2.5% DMSO-containing medium and infected with a single-round luciferase HIV-1 virus. Luciferase activity was detected 24 hours post infection. Depicted are mean values (±SEM) of three biological replicates of one representative experiment out of three. ( f-i ) Cells were infected with HBV inoculum, and where indicated, the entry inhibitor MyrcludexB (MyrB, 200 nM), or the reverse transcription inhibitor lamivudine (Lami, 0,5 μM) were added. Cells were cultured in medium containing 2.5% DMSO. Ten days post infection, HBV DNA from the supernatant ( f ), intracellular total HBV DNA ( g ) and cccDNA ( h ) were quantified by qPCR. ( i ) The intracellular HBV RNAs levels were determined 10 days post infection by RT-qPCR using primer sets that amplify the indicated HBV RNAs (3.5 kb; 3.5 kb and 2.4 kb; 3.5 kb, 2.4 kb and 2.1 kb, or all four HBV RNAs). ( f ) For each individual experiment, fold-changes to the untreated control shRNA were calculated based on the median of three biological replicates, each measured in three technical replicates. The mean ± SEM of the fold-changes of three independent experiments is depicted. ( g – i ) For each independent experiment, fold-changes to untreated control shRNA were calculated based on the median of three technical replicates. The mean ± SEM of the fold-changes of at least 3 independent experiments is represented. ( f – i ) Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Article Snippet: For HBV total DNA, qPCR reactions were performed on undigested samples in SYBR green supermix (Bio-rad 172-5121) using HBV specific primers HBV-total-DNA-F and HBV-total-DNA-R (95 °C for 10 minutes, followed up by 95 °C for 10 seconds and 60 °C for 30 seconds for 40 cycles).

    Techniques: Infection, Western Blot, shRNA, Cell Culture, Expressing, Luciferase, Activity Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Silencing of SAMHD1 increases HBV replication in resting HepG2.2.15 cells. ( a ) SAMHD1 or actin protein levels were detected in cell lysates from SAMHD1 knockdown or control HepG2.2.15 cells by Western blotting. ( b ) Phosphorylation on residue T592 of SAMHD1 and cyclin B1 expression were detected by Western blotting of lysates from control shRNA HepG2.2.15 cells cultured for 3 days in complete versus serum-free medium. ( c-e ) HepG2.2.15 cells stably expressing two different SAMHD1 shRNAs or a control shRNA were cultured in serum-free medium for 3 days. ( c ) Luciferase activity (relative luciferase units, RLU) was detected in cells 24 hours post infection with a single-round HIV-1-based lentiviral vector that encoded luciferase. ( d ) Equal cell growth and viability between different shRNA cell lines was assessed using ATPLite. ( e ) The amount of HBV DNA in the supernatant was determined by qPCR and normalized to the control shRNA. In ( c , d ), the mean ± standard error mean (SEM) of three biological replicates of one representative experiment is depicted. In ( e ), the fold-changes to negative control were calculated for each individual experiment based on the median of three biological replicates, each measured in three technical replicates. The mean ± SEM of the fold-changes of four independent experiments is depicted. Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Journal: Scientific Reports

    Article Title: Restrictive influence of SAMHD1 on Hepatitis B Virus life cycle

    doi: 10.1038/srep26616

    Figure Lengend Snippet: Silencing of SAMHD1 increases HBV replication in resting HepG2.2.15 cells. ( a ) SAMHD1 or actin protein levels were detected in cell lysates from SAMHD1 knockdown or control HepG2.2.15 cells by Western blotting. ( b ) Phosphorylation on residue T592 of SAMHD1 and cyclin B1 expression were detected by Western blotting of lysates from control shRNA HepG2.2.15 cells cultured for 3 days in complete versus serum-free medium. ( c-e ) HepG2.2.15 cells stably expressing two different SAMHD1 shRNAs or a control shRNA were cultured in serum-free medium for 3 days. ( c ) Luciferase activity (relative luciferase units, RLU) was detected in cells 24 hours post infection with a single-round HIV-1-based lentiviral vector that encoded luciferase. ( d ) Equal cell growth and viability between different shRNA cell lines was assessed using ATPLite. ( e ) The amount of HBV DNA in the supernatant was determined by qPCR and normalized to the control shRNA. In ( c , d ), the mean ± standard error mean (SEM) of three biological replicates of one representative experiment is depicted. In ( e ), the fold-changes to negative control were calculated for each individual experiment based on the median of three biological replicates, each measured in three technical replicates. The mean ± SEM of the fold-changes of four independent experiments is depicted. Statistical significance was determined using a one-way ANOVA with multiple comparisons according to Dunnett (*p

    Article Snippet: For HBV total DNA, qPCR reactions were performed on undigested samples in SYBR green supermix (Bio-rad 172-5121) using HBV specific primers HBV-total-DNA-F and HBV-total-DNA-R (95 °C for 10 minutes, followed up by 95 °C for 10 seconds and 60 °C for 30 seconds for 40 cycles).

    Techniques: Western Blot, Expressing, shRNA, Cell Culture, Stable Transfection, Luciferase, Activity Assay, Infection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Negative Control

    Cloned PCR products from the 5′ (exon 4, italic) and 3′ (exon 16, bold) primer (squared) combination of CD44 in A2058 human melanoma cell line. Direct sequencing shows a CD44 isoform with no v1 or any other variable exons (A) as well as one with truncated v1 (underlined).

    Journal: PLoS ONE

    Article Title: Demonstration of a Melanoma-Specific CD44 Alternative Splicing Pattern That Remains Qualitatively Stable, but Shows Quantitative Changes during Tumour Progression

    doi: 10.1371/journal.pone.0053883

    Figure Lengend Snippet: Cloned PCR products from the 5′ (exon 4, italic) and 3′ (exon 16, bold) primer (squared) combination of CD44 in A2058 human melanoma cell line. Direct sequencing shows a CD44 isoform with no v1 or any other variable exons (A) as well as one with truncated v1 (underlined).

    Article Snippet: Quantitative PCR Analysis For quantitative measurement of the expressed CD44 variable exons q-PCR reactions were used (iQ SYBR® Green Supermix,Bio-Rad), by cycling conditions 3 min at 95°C, then 40 cycles at 95°C 30 sec, 55°C 30 sec, 72°C 1 min.

    Techniques: Clone Assay, Polymerase Chain Reaction, Sequencing

    Relative quantitative expression of CD44 variable exons in cell cultures from metastatic (newborn) and non-metastatic [adult (AP)] human xenograft model (Real-Time PCR measurement) of HT168M1, a human melanoma cell line of originally high variable exon expression level. A. The relative expression level of all variable exons is raised in certain lung metastasis (NM), when remaines low or even decreases in other lung metastases resulting in large error bars. It should be noted that the expression level of the primaries from different localisations [newborn primary (NP), adult primary (AP) and intravenously implanted lung colony (IVLC)] are all comparable, although the slightly higher expression observed in the adult primary is an unexpected finding. B. We used the lung metastasis from the newborn animal with the highest CD44 variable exon expression level (NM = S1T2) for subcutaneous re-implantation into another newborn animal. The expression level in the primary tumour (PNM) and its metastases (MPNM) was 24 times lower on average. C. The liver metastases (LMIVLC) from the intravenously implanted lung colonies (IVLC) showed a decrease in expression, when compared to the lung colonies (IVLC).

    Journal: PLoS ONE

    Article Title: Demonstration of a Melanoma-Specific CD44 Alternative Splicing Pattern That Remains Qualitatively Stable, but Shows Quantitative Changes during Tumour Progression

    doi: 10.1371/journal.pone.0053883

    Figure Lengend Snippet: Relative quantitative expression of CD44 variable exons in cell cultures from metastatic (newborn) and non-metastatic [adult (AP)] human xenograft model (Real-Time PCR measurement) of HT168M1, a human melanoma cell line of originally high variable exon expression level. A. The relative expression level of all variable exons is raised in certain lung metastasis (NM), when remaines low or even decreases in other lung metastases resulting in large error bars. It should be noted that the expression level of the primaries from different localisations [newborn primary (NP), adult primary (AP) and intravenously implanted lung colony (IVLC)] are all comparable, although the slightly higher expression observed in the adult primary is an unexpected finding. B. We used the lung metastasis from the newborn animal with the highest CD44 variable exon expression level (NM = S1T2) for subcutaneous re-implantation into another newborn animal. The expression level in the primary tumour (PNM) and its metastases (MPNM) was 24 times lower on average. C. The liver metastases (LMIVLC) from the intravenously implanted lung colonies (IVLC) showed a decrease in expression, when compared to the lung colonies (IVLC).

    Article Snippet: Quantitative PCR Analysis For quantitative measurement of the expressed CD44 variable exons q-PCR reactions were used (iQ SYBR® Green Supermix,Bio-Rad), by cycling conditions 3 min at 95°C, then 40 cycles at 95°C 30 sec, 55°C 30 sec, 72°C 1 min.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Relative quantitative expression of CD44 variable exons in cell cultures from metastatic (newborn) and non-metastatic human xenograft model (Real-Time PCR measurement) of HT199, a human melanoma cell line of originally low variable exon expression level. A. The relative expression level of all variable exons is raised in circulating metastatic cells (NCTC) and metastatic cells (NM) compared to their levels in primary tumours [newborn primary (NP) and adult primary (AP)] and lung colony (IVLC) B. The qualitative fingerprint (bottom line) remains unchanged.

    Journal: PLoS ONE

    Article Title: Demonstration of a Melanoma-Specific CD44 Alternative Splicing Pattern That Remains Qualitatively Stable, but Shows Quantitative Changes during Tumour Progression

    doi: 10.1371/journal.pone.0053883

    Figure Lengend Snippet: Relative quantitative expression of CD44 variable exons in cell cultures from metastatic (newborn) and non-metastatic human xenograft model (Real-Time PCR measurement) of HT199, a human melanoma cell line of originally low variable exon expression level. A. The relative expression level of all variable exons is raised in circulating metastatic cells (NCTC) and metastatic cells (NM) compared to their levels in primary tumours [newborn primary (NP) and adult primary (AP)] and lung colony (IVLC) B. The qualitative fingerprint (bottom line) remains unchanged.

    Article Snippet: Quantitative PCR Analysis For quantitative measurement of the expressed CD44 variable exons q-PCR reactions were used (iQ SYBR® Green Supermix,Bio-Rad), by cycling conditions 3 min at 95°C, then 40 cycles at 95°C 30 sec, 55°C 30 sec, 72°C 1 min.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Expression profiling of rice tetraspanin genes in different tissues of rice. (A) Quantitative PCR analysis of transcript levels of rice tetraspanins in tissues namely shoot, root, young leaf (YL), active tillering phase (AT-Phase), stem elongation phase (SE-Phase), spikelets (Spks), young flag leaf (YFL), and mature flag leaf (MFL). For root tissue normalized fold change (log 2 scale) was calculated relative to that in shoot tissue of 7-day-old seedlings. For tissues obtained from field grown plants fold change (log 2 scale) was calculated relative to that in young leaf. (B) Quantitative PCR analysis of transcript levels of rice tetraspanins during progression of senescence in flag leaf of field-grown rice plants. CON: fully expanded MFL; 100% chlorophyll, S1: 80–90% chlorophyll; S2: 60–80% chlorophyll; S3: 40–60% chlorophyll. Normalized fold change (log 2 scale) was calculated relative to that in MFL. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Error bars represent standard error (SE) of three independent biological replicates.

    Journal: Frontiers in Plant Science

    Article Title: Comprehensive Expression Profiling of Rice Tetraspanin Genes Reveals Diverse Roles During Development and Abiotic Stress

    doi: 10.3389/fpls.2015.01088

    Figure Lengend Snippet: Expression profiling of rice tetraspanin genes in different tissues of rice. (A) Quantitative PCR analysis of transcript levels of rice tetraspanins in tissues namely shoot, root, young leaf (YL), active tillering phase (AT-Phase), stem elongation phase (SE-Phase), spikelets (Spks), young flag leaf (YFL), and mature flag leaf (MFL). For root tissue normalized fold change (log 2 scale) was calculated relative to that in shoot tissue of 7-day-old seedlings. For tissues obtained from field grown plants fold change (log 2 scale) was calculated relative to that in young leaf. (B) Quantitative PCR analysis of transcript levels of rice tetraspanins during progression of senescence in flag leaf of field-grown rice plants. CON: fully expanded MFL; 100% chlorophyll, S1: 80–90% chlorophyll; S2: 60–80% chlorophyll; S3: 40–60% chlorophyll. Normalized fold change (log 2 scale) was calculated relative to that in MFL. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Error bars represent standard error (SE) of three independent biological replicates.

    Article Snippet: Quantitative PCR (qPCR) was performed with three biological replicates and two technical replicates using Sso fast Evagreen supermix (Bio-Rad, USA), appropriate primers and Master cycler RealPlex2 (Eppendorf, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to nutrient deprivation. Seven-day-old rice seedlings were grown in different nutrient deprivation media such as nitrogen deprivation (N), phosphorous deprivation (P), potassium deprivation (K), sulfur deprivation (S) for different time durations as indicated on the left (in h). Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in unstressed seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Journal: Frontiers in Plant Science

    Article Title: Comprehensive Expression Profiling of Rice Tetraspanin Genes Reveals Diverse Roles During Development and Abiotic Stress

    doi: 10.3389/fpls.2015.01088

    Figure Lengend Snippet: Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to nutrient deprivation. Seven-day-old rice seedlings were grown in different nutrient deprivation media such as nitrogen deprivation (N), phosphorous deprivation (P), potassium deprivation (K), sulfur deprivation (S) for different time durations as indicated on the left (in h). Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in unstressed seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Article Snippet: Quantitative PCR (qPCR) was performed with three biological replicates and two technical replicates using Sso fast Evagreen supermix (Bio-Rad, USA), appropriate primers and Master cycler RealPlex2 (Eppendorf, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Software

    Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to different abiotic stresses. Seven-day-old rice seedlings were exposed to different abiotic stresses such as heat stress at 42°C; salinity stress with 200 mM NaCl; water deficit stress (WDS) imposed by 15% PEG; cold stress at 4°C; oxidative stress with 10 mM H 2 O 2 for different time durations as indicated on the left (duration in h). Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in unstressed seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Journal: Frontiers in Plant Science

    Article Title: Comprehensive Expression Profiling of Rice Tetraspanin Genes Reveals Diverse Roles During Development and Abiotic Stress

    doi: 10.3389/fpls.2015.01088

    Figure Lengend Snippet: Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to different abiotic stresses. Seven-day-old rice seedlings were exposed to different abiotic stresses such as heat stress at 42°C; salinity stress with 200 mM NaCl; water deficit stress (WDS) imposed by 15% PEG; cold stress at 4°C; oxidative stress with 10 mM H 2 O 2 for different time durations as indicated on the left (duration in h). Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in unstressed seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Article Snippet: Quantitative PCR (qPCR) was performed with three biological replicates and two technical replicates using Sso fast Evagreen supermix (Bio-Rad, USA), appropriate primers and Master cycler RealPlex2 (Eppendorf, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Software

    Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to different hormones. Seven-day-old seedlings were exposed to exogenous hormones namely abscisic acid (ABA), brassinosteroids (BS), gibberellic acid (GA), and methyl jasmonate (MeJA) for different time durations (in h) as indicated on the left. Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in untreated seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Journal: Frontiers in Plant Science

    Article Title: Comprehensive Expression Profiling of Rice Tetraspanin Genes Reveals Diverse Roles During Development and Abiotic Stress

    doi: 10.3389/fpls.2015.01088

    Figure Lengend Snippet: Heat map representing expression profile of rice tetraspanin genes in rice seedlings exposed to different hormones. Seven-day-old seedlings were exposed to exogenous hormones namely abscisic acid (ABA), brassinosteroids (BS), gibberellic acid (GA), and methyl jasmonate (MeJA) for different time durations (in h) as indicated on the left. Expression levels of tetraspanin genes were determined by quantitative PCR and normalized fold change (log 2 scale) was calculated relative to that in untreated seedlings. For normalization eEF-1α was used as internal control. Three biological replicates and two technical replicates were included in the study. Hierarchical clustering analysis of relative fold change was performed to prepare a dendrogram and a heat map using Hierarchical Clustering Explorer v3.5 software.

    Article Snippet: Quantitative PCR (qPCR) was performed with three biological replicates and two technical replicates using Sso fast Evagreen supermix (Bio-Rad, USA), appropriate primers and Master cycler RealPlex2 (Eppendorf, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Software

    IL-17A does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time PCR analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.

    Journal: The Journal of allergy and clinical immunology

    Article Title: IL-17A enhances IL-13 activity by enhancing IL-13-induced STAT6 activation

    doi: 10.1016/j.jaci.2016.04.037

    Figure Lengend Snippet: IL-17A does not stabilize IL-13-induced transcripts. Percentage mRNA remaining of IL-13 induced C3 ( A ) and Il13ra2 ( B ) in NIH/3T3s stimulated with IL-13 or IL-13 + IL-17A for 16 hours and incubated with ActD. Real-time PCR analysis of transcript levels was evaluated over 24 hours and t½ was calculated. Mean + SEM of 4-8 replicates per condition shown. Results show 1 of 3 independent experiments.

    Article Snippet: To assess IL-13 and IL-17A-induced message expression in lung sections and cell cultures, we used quantitative real-time PCR with SYBR Green mix (Biorad).

    Techniques: Incubation, Real-time Polymerase Chain Reaction

    Reciprocal co-regulation of IL-13- and IL-17A-induced genes occurs in vitro. Real-time PCR analysis of C3 ( A ), Il13ra2 ( B ), Cebpd ( C ), and Lcn2 ( D ) expression in NIH/3T3s stimulated with IL-13, IL-17A, or both cytokines over 24 hours. ## and ### indicate P

    Journal: The Journal of allergy and clinical immunology

    Article Title: IL-17A enhances IL-13 activity by enhancing IL-13-induced STAT6 activation

    doi: 10.1016/j.jaci.2016.04.037

    Figure Lengend Snippet: Reciprocal co-regulation of IL-13- and IL-17A-induced genes occurs in vitro. Real-time PCR analysis of C3 ( A ), Il13ra2 ( B ), Cebpd ( C ), and Lcn2 ( D ) expression in NIH/3T3s stimulated with IL-13, IL-17A, or both cytokines over 24 hours. ## and ### indicate P

    Article Snippet: To assess IL-13 and IL-17A-induced message expression in lung sections and cell cultures, we used quantitative real-time PCR with SYBR Green mix (Biorad).

    Techniques: In Vitro, Real-time Polymerase Chain Reaction, Expressing

    IL-17A directly signals on IL-13-responsive cells to enhance gene expression. Real-time PCR analysis of C3 ( A ), Il13ra2 ( B ), and Lcn2 ( C ) expression in co-cultured WT or IL-17RA −/− lung fibroblasts stimulated with IL-13, IL-17A, or both cytokines for 24 hours. # and ### indicate P

    Journal: The Journal of allergy and clinical immunology

    Article Title: IL-17A enhances IL-13 activity by enhancing IL-13-induced STAT6 activation

    doi: 10.1016/j.jaci.2016.04.037

    Figure Lengend Snippet: IL-17A directly signals on IL-13-responsive cells to enhance gene expression. Real-time PCR analysis of C3 ( A ), Il13ra2 ( B ), and Lcn2 ( C ) expression in co-cultured WT or IL-17RA −/− lung fibroblasts stimulated with IL-13, IL-17A, or both cytokines for 24 hours. # and ### indicate P

    Article Snippet: To assess IL-13 and IL-17A-induced message expression in lung sections and cell cultures, we used quantitative real-time PCR with SYBR Green mix (Biorad).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Cell Culture

    IL-17A directly enhances IL-13-driven pSTAT6. pSTAT6/STAT6 levels in NIH/3T3 cells pretreated with DMSO ( A ) or cycloheximide ( B ) then stimulated with cytokines for 5 minutes, ( C ) densitometry of pSTAT6/STAT6 expression shown in ( A , B ), geometric mean fluorescence intensity of pSTAT6 by phosphoflow in NIH/3T3s stimulated with cytokines over 1 hour ( D ), pSTAT6 and STAT6 expression and associated densitometry in whole lung homogenates from cytokine-treated A/J mice (mean + SEM of n=7-10 mice per group, pooled from 3 independent experiments) ( E ), real-time PCR analysis of C3 and Il13ra2 ( F ) or Lcn2 ( G ) expression in WT and STAT6 −/− lung fibroblasts stimulated with cytokines for 24 hours. #, ##, and ### indicate P

    Journal: The Journal of allergy and clinical immunology

    Article Title: IL-17A enhances IL-13 activity by enhancing IL-13-induced STAT6 activation

    doi: 10.1016/j.jaci.2016.04.037

    Figure Lengend Snippet: IL-17A directly enhances IL-13-driven pSTAT6. pSTAT6/STAT6 levels in NIH/3T3 cells pretreated with DMSO ( A ) or cycloheximide ( B ) then stimulated with cytokines for 5 minutes, ( C ) densitometry of pSTAT6/STAT6 expression shown in ( A , B ), geometric mean fluorescence intensity of pSTAT6 by phosphoflow in NIH/3T3s stimulated with cytokines over 1 hour ( D ), pSTAT6 and STAT6 expression and associated densitometry in whole lung homogenates from cytokine-treated A/J mice (mean + SEM of n=7-10 mice per group, pooled from 3 independent experiments) ( E ), real-time PCR analysis of C3 and Il13ra2 ( F ) or Lcn2 ( G ) expression in WT and STAT6 −/− lung fibroblasts stimulated with cytokines for 24 hours. #, ##, and ### indicate P

    Article Snippet: To assess IL-13 and IL-17A-induced message expression in lung sections and cell cultures, we used quantitative real-time PCR with SYBR Green mix (Biorad).

    Techniques: Expressing, Fluorescence, Mouse Assay, Real-time Polymerase Chain Reaction

    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. Reverse transcription–polymerase chain reaction showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. Reverse transcription–polymerase chain reaction showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

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

    SB rapidly induces progressed phenotype in epidermis-expressing human C-Ha RASV12G oncogene. ( A ) Significant increase in proliferation with SB treatment in RAS -expressing skin (left) and slight increase in epidermal thickness (right). ( B ) An increase in MPO+ cells in the dermis of SB-treated mice expressing the InvtTA × tetO RASV12G transgenes. Arrows indicate neutrophils. Yellow line shows epidermal/dermal junction. Representative micrographs (right), magnification, ×100. Scale bar represents 50 μm. ( C ) Increased messenger RNA expression of neutrophil chemokines s100a8, s100a9 and keratinocyte chemokine with treatment of SB. ( D ) SB causes significant decrease in keratin 1 staining (left) and messenger RNA expression (middle). Representative micrographs (left), magnification ×100. Scale bar represents 50 μm. Yellow dotted line shows epidermal/dermal junction. Significant increase in keratin 18 messenger RNA expression by quantitative reverse transcription–polymerase chain reaction in RAS -expressing skin following SB (right). n = 4–6 mice per treatment group.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB rapidly induces progressed phenotype in epidermis-expressing human C-Ha RASV12G oncogene. ( A ) Significant increase in proliferation with SB treatment in RAS -expressing skin (left) and slight increase in epidermal thickness (right). ( B ) An increase in MPO+ cells in the dermis of SB-treated mice expressing the InvtTA × tetO RASV12G transgenes. Arrows indicate neutrophils. Yellow line shows epidermal/dermal junction. Representative micrographs (right), magnification, ×100. Scale bar represents 50 μm. ( C ) Increased messenger RNA expression of neutrophil chemokines s100a8, s100a9 and keratinocyte chemokine with treatment of SB. ( D ) SB causes significant decrease in keratin 1 staining (left) and messenger RNA expression (middle). Representative micrographs (left), magnification ×100. Scale bar represents 50 μm. Yellow dotted line shows epidermal/dermal junction. Significant increase in keratin 18 messenger RNA expression by quantitative reverse transcription–polymerase chain reaction in RAS -expressing skin following SB (right). n = 4–6 mice per treatment group.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

    Techniques: Expressing, Mouse Assay, RNA Expression, Staining, Reverse Transcription Polymerase Chain Reaction

    SB reduces TPA-induced Smad2 phosphorylation in vivo . ( A (available at Carcinogenesis Online). ( B ) Quantitative reverse transcription–polymerase chain reaction analysis of Smad7 and LTBP-1 gene expression in acetone or SB-treated whole skin. Normalized to acetone only samples ( n = 4). ( C ) Indirect immunofluorescence showing a reduction in TPA-induced p-Smad2 by SB at 4 and 24 h TPA treatment. Magnification ×800, bar represents 15 μm. P-Smad2 was detected with Alexa-fluor-488 (green), and nuclei counterstained with TO-PRO3 (red), (representative of n (available at Carcinogenesis Online). ( D ) Western blot of whole skin treated 24 h with TPA and 12 h SB, showing reduced TPA-induced Smad2 phosphorylation with SB treatment (representative of n (available at Carcinogenesis Online). C, control and A, acetone.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB reduces TPA-induced Smad2 phosphorylation in vivo . ( A (available at Carcinogenesis Online). ( B ) Quantitative reverse transcription–polymerase chain reaction analysis of Smad7 and LTBP-1 gene expression in acetone or SB-treated whole skin. Normalized to acetone only samples ( n = 4). ( C ) Indirect immunofluorescence showing a reduction in TPA-induced p-Smad2 by SB at 4 and 24 h TPA treatment. Magnification ×800, bar represents 15 μm. P-Smad2 was detected with Alexa-fluor-488 (green), and nuclei counterstained with TO-PRO3 (red), (representative of n (available at Carcinogenesis Online). ( D ) Western blot of whole skin treated 24 h with TPA and 12 h SB, showing reduced TPA-induced Smad2 phosphorylation with SB treatment (representative of n (available at Carcinogenesis Online). C, control and A, acetone.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

    Techniques: In Vivo, Reverse Transcription Polymerase Chain Reaction, Expressing, Immunofluorescence, Western Blot

    (A) SYBR green dissociation curve analyses of LSG-qPCR amplicons from the Stratagene Mx3000P platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis

    Journal: Journal of Clinical Microbiology

    Article Title: Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay

    doi: 10.1128/JCM.01764-16

    Figure Lengend Snippet: (A) SYBR green dissociation curve analyses of LSG-qPCR amplicons from the Stratagene Mx3000P platform. G1, L . ( V .) braziliensis , L . ( V .) guyanensis , and L . ( V .) panamensis ; G2A/B, L . ( L .) infantum/L . ( L .) chagasi and L . ( L .) tropica ; G3, L . ( L .) amazonensis

    Article Snippet: PCRs were run on either of two different qPCR platforms—the Stratagene Mx3000P qPCR system (Stratagene-Agilent) or the ABI 7500 real-time PCR system (ABI Life Technologies)—using the following cycling parameters: 95°C for 15 min and 40 cycles of 95°C for 15 s and 60°C for 1 min. Fluorescence data were collected at the end of each 60°C plateau.

    Techniques: SYBR Green Assay, Real-time Polymerase Chain Reaction