for qrt pcr Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Thermo Fisher transcript polymerase chain reaction qrt pcr kit
    Transcript Polymerase Chain Reaction Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transcript polymerase chain reaction qrt pcr kit/product/Thermo Fisher
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    transcript polymerase chain reaction qrt pcr kit - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    91
    TaKaRa transcription polymerase chain reaction qrt pcr system
    HIF-1α regulated the expression of CCL2 in PDAC cells, PC, N and H indicate positive control, normoxia, and hypoxia, respectively. ( A ) HPDE6-C7,BxPC3 and MiaPaca-2 cells were transfected with siHIF-1α (50 nmol/L) and pcDNA3.1-HIF-1α plasmids (4 µg) for 48 h, respectively; the mRNA expression levels of HIF-1α and CCL2 were assessed by <t>qRT-PCR.</t> The experiments were performed thrice independently. * p
    Transcription Polymerase Chain Reaction Qrt Pcr System, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transcription polymerase chain reaction qrt pcr system/product/TaKaRa
    Average 91 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    transcription polymerase chain reaction qrt pcr system - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    80
    Thermo Fisher polymerase chain reaction qrt pcr rneasy mini kit
    HIF-1α regulated the expression of CCL2 in PDAC cells, PC, N and H indicate positive control, normoxia, and hypoxia, respectively. ( A ) HPDE6-C7,BxPC3 and MiaPaca-2 cells were transfected with siHIF-1α (50 nmol/L) and pcDNA3.1-HIF-1α plasmids (4 µg) for 48 h, respectively; the mRNA expression levels of HIF-1α and CCL2 were assessed by <t>qRT-PCR.</t> The experiments were performed thrice independently. * p
    Polymerase Chain Reaction Qrt Pcr Rneasy Mini Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr rneasy mini kit/product/Thermo Fisher
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr rneasy mini kit - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

    99
    Thermo Fisher real time polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1001 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 99 stars, based on 1001 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    97
    Thermo Fisher reverse transcription polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 540 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcription polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 97 stars, based on 540 article reviews
    Price from $9.99 to $1999.99
    reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    91
    Thermo Fisher reverse transcriptase polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcriptase polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    reverse transcriptase polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    99
    TaKaRa real time polymerase chain reaction qrt pcr
    The effect of TGF-β3 by TCDD induced in MEPM cells. 0.5 nM, 1 nM, 5 nM, 10 nM, 20 nM and 50 nM TCDD, or DMSO (≤0.05%)-treated MEPM cells as the experiment group and control group, respectively. After treated for 72 hours, the expression of TGF-β3 was measured by <t>qRT-PCR.</t> Data were mean values ± standard deviation of 3 replicate experiments. * P
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 231 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr/product/TaKaRa
    Average 99 stars, based on 231 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    92
    Bio-Rad polymerase chain reaction qrt pcr
    CDKI expression in human terminal erythroid differentiation. (A) RNA-seq data showing CDKI members’ expression (fragments per kilobase of transcript per million) at each distinct stage of human terminal erythroid differentiation. Baso-E, basophilic erythroblast; ortho-E, orthochromatic erythroblast; poly-E, polychromatic erythroblast; pro-E, proerythroblast. (B) Representative image of western blotting of p19 INK4d , p18 INK4c , and p27 KIP1 expression in whole-cell lysates prepared from erythroblasts cultured for different times (left). Quantitative analysis of data from 3 independent experiments of protein expression levels (right). GAPDH was used as a loading control. (C) <t>qRT-PCR</t> results showing p19 INK4d expression in erythroblasts infected with lentivirus containing control (Lucif shRNA) or p19 INK4d shRNA on day (D) 7, D 11, and D 15 of culture. The results were normalized to GAPDH mRNA. (D) Representative images of western blotting showing p19 INK4d expression levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (left). Quantitative analysis of protein expression data from 3 independent experiments (right). The results were normalized to GAPDH protein. Statistical analysis of data of 3 independent experiments and bar plot represents mean ± SD of triplicate samples. *** P
    Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr/product/Bio-Rad
    Average 92 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    88
    Roche polymerase chain reaction qrt pcr
    CDKI expression in human terminal erythroid differentiation. (A) RNA-seq data showing CDKI members’ expression (fragments per kilobase of transcript per million) at each distinct stage of human terminal erythroid differentiation. Baso-E, basophilic erythroblast; ortho-E, orthochromatic erythroblast; poly-E, polychromatic erythroblast; pro-E, proerythroblast. (B) Representative image of western blotting of p19 INK4d , p18 INK4c , and p27 KIP1 expression in whole-cell lysates prepared from erythroblasts cultured for different times (left). Quantitative analysis of data from 3 independent experiments of protein expression levels (right). GAPDH was used as a loading control. (C) <t>qRT-PCR</t> results showing p19 INK4d expression in erythroblasts infected with lentivirus containing control (Lucif shRNA) or p19 INK4d shRNA on day (D) 7, D 11, and D 15 of culture. The results were normalized to GAPDH mRNA. (D) Representative images of western blotting showing p19 INK4d expression levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (left). Quantitative analysis of protein expression data from 3 independent experiments (right). The results were normalized to GAPDH protein. Statistical analysis of data of 3 independent experiments and bar plot represents mean ± SD of triplicate samples. *** P
    Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr/product/Roche
    Average 88 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    88/100 stars
      Buy from Supplier

    86
    Thermo Fisher qrt polymerase chain reaction pcr
    CDKI expression in human terminal erythroid differentiation. (A) RNA-seq data showing CDKI members’ expression (fragments per kilobase of transcript per million) at each distinct stage of human terminal erythroid differentiation. Baso-E, basophilic erythroblast; ortho-E, orthochromatic erythroblast; poly-E, polychromatic erythroblast; pro-E, proerythroblast. (B) Representative image of western blotting of p19 INK4d , p18 INK4c , and p27 KIP1 expression in whole-cell lysates prepared from erythroblasts cultured for different times (left). Quantitative analysis of data from 3 independent experiments of protein expression levels (right). GAPDH was used as a loading control. (C) <t>qRT-PCR</t> results showing p19 INK4d expression in erythroblasts infected with lentivirus containing control (Lucif shRNA) or p19 INK4d shRNA on day (D) 7, D 11, and D 15 of culture. The results were normalized to GAPDH mRNA. (D) Representative images of western blotting showing p19 INK4d expression levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (left). Quantitative analysis of protein expression data from 3 independent experiments (right). The results were normalized to GAPDH protein. Statistical analysis of data of 3 independent experiments and bar plot represents mean ± SD of triplicate samples. *** P
    Qrt Polymerase Chain Reaction Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt polymerase chain reaction pcr/product/Thermo Fisher
    Average 86 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    qrt polymerase chain reaction pcr - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    98
    Roche real time polymerase chain reaction qrt pcr
    LPS / ATP activate NLRP 3 inflammasomes in VSMC s. Human primary VSMC s were treated with vehicle or the indicated concentrations of LPS for 16 hours and/or ATP for 1 hour. Some cells were transfected with control or NLRP 3‐specific si RNA or treated with vehicle DMSO or ZYVAD ‐ FMK , followed by treatment with LPS and/or ATP . The relative levels of NLRP 3 and cleaved caspase‐1 expression were determined by <t>qRT</t> ‐ <t>PCR</t> and Western blot. The distribution of HMGB 1 in the different groups of cells and their cultured supernatants were determined by Western blot and ELISA . Data are representative images or expressed as the mean± SD of each group from 4 separated experiments (Western blot) or 3 separated experiments with 4 duplicated wells ( qRT ‐ PCR and ELISA ). A, The levels of NLRP 3 mRNA transcripts. B, The levels of NLRP 3 proteins. C, The levels of cleaved caspase‐1. D, The levels of total HMGB 1. E, The levels of nuclear HMGB 1. F, The levels of cytoplasmic HMGB 1. G, The levels of HMGB 1 in the supernatants. * P
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 98/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr/product/Roche
    Average 98 stars, based on 168 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    98/100 stars
      Buy from Supplier

    97
    Bio-Rad real time polymerase chain reaction qrt pcr
    Elevated levels of Mg 2+ in APP/PS1 transgenic mice decrease the expression of IL-1β. The APP/PS1 transgenic mice at the age of 4 months were administered Mg 2+ (100 mg/kg/d) for 2 months before collecting the brain ( a ). In select experiments, the brains of APP/PS1 transgenic mice at the age of 3 months were harvested and sectioned (400 μm) using a cryostat ( b ). In separate experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) and the right cerebral ventricle was injected (i.c.v.) with D1A cells, which was pre-transfected with IL-1β promoter in the right cerebral ventricle ( c ). In distinct experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) before staining with IL-1β antibody and scanning under two-photon microscopy ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody (a left panel, b). These images are representative of six independent experiments, all with similar results. IL-1β protein and mRNA levels were determined by <t>qRT-PCR</t> and IL-1β enzyme immunoassay kits, respectively (a right panel). The total amounts of GAPDH and protein served as an internal control. The experimental cartoon and real surgery images are shown (c, d upper panel). Luciferase activities from the different groups of mice were measured using a live animal imaging system (c lower panel). The immunofluorescence of IL-1β was scanned using a two-photon microscope (d lower panel). The data represent the means ± S.E. of three independent experiments. * p
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr/product/Bio-Rad
    Average 97 stars, based on 245 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    99
    Bio-Rad quantitative real time polymerase chain reaction qrt pcr
    The preventive effects of AMPK on the senescence associated with NAD + synthesis. (A) Cellular concentrations of NAD + on day 3 or day 5 after H 2 O 2 treatment. (B) Representative images of SA ‐β‐Gal staining of cells (left) and percentage of SA ‐β‐Gal‐positive cells (right). (C) Cellular concentrations of NAD + in the cells incubated with nicotinamide mononucleotide ( NMN ) for 3 days after H 2 O 2 treatment. (D) H 2 O 2 ‐treated cells were incubated with Met (10 mM ), BBR (10 μM), or CC (10 μM) for 3 days. The concentrations of NAD + are shown. (E) A schematic diagram of two pathways of NAD + synthesis. Red ones indicate rate‐limiting enzymes, and blue ones illustrate the enzyme inhibitors we used. (F) Relative fold‐changes in the mRNA levels of NAMPT , as monitored by <t>qRT</t> ‐ <t>PCR</t> (left), and representative images from immunoblot assays against NAMPT are shown (right). The ratio of NAMPT to actin was quantified by densitometry, based on the immunoblot images from three independent experiments. (G) Cells were incubated with Met, BBR , and CC for 3 days. Representative images from immunoblots against NAMPT are shown. The ratio of NAMPT to β‐actin was quantified by densitometry based on the immunoblot images from three independent experiments. (H) H 2 O 2 ‐treated NIH 3T3 cells were incubated with NAMPT inhibitor ( FK 866, 5 nM ), QPRT inhibitor ( PHTH , 1 mM ), Met (10 mM ), and BBR (10 μM) alone or in combination as indicated for 3 days. The percentages of SA ‐β‐Gal‐positive cells are shown. (I) The mRNA levels of NMNAT 1 , NMNAT 2 , and NMNAT 3 genes in the cells infected with corresponding lentivirus‐sh RNA or nontargeting sh RNA (sh CON ) as determined by qRT ‐ PCR . (J) Cellular concentrations of NAD + in sh RNA ‐infected cells. (K) Cells infected with sh RNA s were treated with H 2 O 2 and incubated with BBR for 3 days. Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). * P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 561 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr/product/Bio-Rad
    Average 99 stars, based on 561 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    94
    Agilent technologies quantitative real time polymerase chain reaction qrt pcr
    The preventive effects of AMPK on the senescence associated with NAD + synthesis. (A) Cellular concentrations of NAD + on day 3 or day 5 after H 2 O 2 treatment. (B) Representative images of SA ‐β‐Gal staining of cells (left) and percentage of SA ‐β‐Gal‐positive cells (right). (C) Cellular concentrations of NAD + in the cells incubated with nicotinamide mononucleotide ( NMN ) for 3 days after H 2 O 2 treatment. (D) H 2 O 2 ‐treated cells were incubated with Met (10 mM ), BBR (10 μM), or CC (10 μM) for 3 days. The concentrations of NAD + are shown. (E) A schematic diagram of two pathways of NAD + synthesis. Red ones indicate rate‐limiting enzymes, and blue ones illustrate the enzyme inhibitors we used. (F) Relative fold‐changes in the mRNA levels of NAMPT , as monitored by <t>qRT</t> ‐ <t>PCR</t> (left), and representative images from immunoblot assays against NAMPT are shown (right). The ratio of NAMPT to actin was quantified by densitometry, based on the immunoblot images from three independent experiments. (G) Cells were incubated with Met, BBR , and CC for 3 days. Representative images from immunoblots against NAMPT are shown. The ratio of NAMPT to β‐actin was quantified by densitometry based on the immunoblot images from three independent experiments. (H) H 2 O 2 ‐treated NIH 3T3 cells were incubated with NAMPT inhibitor ( FK 866, 5 nM ), QPRT inhibitor ( PHTH , 1 mM ), Met (10 mM ), and BBR (10 μM) alone or in combination as indicated for 3 days. The percentages of SA ‐β‐Gal‐positive cells are shown. (I) The mRNA levels of NMNAT 1 , NMNAT 2 , and NMNAT 3 genes in the cells infected with corresponding lentivirus‐sh RNA or nontargeting sh RNA (sh CON ) as determined by qRT ‐ PCR . (J) Cellular concentrations of NAD + in sh RNA ‐infected cells. (K) Cells infected with sh RNA s were treated with H 2 O 2 and incubated with BBR for 3 days. Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). * P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr/product/Agilent technologies
    Average 94 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    89
    Roche polymerase chain reaction qrt pcr analysis
    The preventive effects of AMPK on the senescence associated with NAD + synthesis. (A) Cellular concentrations of NAD + on day 3 or day 5 after H 2 O 2 treatment. (B) Representative images of SA ‐β‐Gal staining of cells (left) and percentage of SA ‐β‐Gal‐positive cells (right). (C) Cellular concentrations of NAD + in the cells incubated with nicotinamide mononucleotide ( NMN ) for 3 days after H 2 O 2 treatment. (D) H 2 O 2 ‐treated cells were incubated with Met (10 mM ), BBR (10 μM), or CC (10 μM) for 3 days. The concentrations of NAD + are shown. (E) A schematic diagram of two pathways of NAD + synthesis. Red ones indicate rate‐limiting enzymes, and blue ones illustrate the enzyme inhibitors we used. (F) Relative fold‐changes in the mRNA levels of NAMPT , as monitored by <t>qRT</t> ‐ <t>PCR</t> (left), and representative images from immunoblot assays against NAMPT are shown (right). The ratio of NAMPT to actin was quantified by densitometry, based on the immunoblot images from three independent experiments. (G) Cells were incubated with Met, BBR , and CC for 3 days. Representative images from immunoblots against NAMPT are shown. The ratio of NAMPT to β‐actin was quantified by densitometry based on the immunoblot images from three independent experiments. (H) H 2 O 2 ‐treated NIH 3T3 cells were incubated with NAMPT inhibitor ( FK 866, 5 nM ), QPRT inhibitor ( PHTH , 1 mM ), Met (10 mM ), and BBR (10 μM) alone or in combination as indicated for 3 days. The percentages of SA ‐β‐Gal‐positive cells are shown. (I) The mRNA levels of NMNAT 1 , NMNAT 2 , and NMNAT 3 genes in the cells infected with corresponding lentivirus‐sh RNA or nontargeting sh RNA (sh CON ) as determined by qRT ‐ PCR . (J) Cellular concentrations of NAD + in sh RNA ‐infected cells. (K) Cells infected with sh RNA s were treated with H 2 O 2 and incubated with BBR for 3 days. Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). * P
    Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr analysis/product/Roche
    Average 89 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-04
    89/100 stars
      Buy from Supplier

    93
    TaKaRa realtime polymerase chain reaction qrt pcr
    <t>qRT-PCR</t> analysis of IL-17 and forkhead box P3 (Foxp3) in the spleen ( n = 3). Note that the Foxp3 level was increased, but the IL-17 level was similar in the spleen of the CAN group compared with the NC group. *** p
    Realtime Polymerase Chain Reaction Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/realtime polymerase chain reaction qrt pcr/product/TaKaRa
    Average 93 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    realtime polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    95
    Genewiz reverse transcription polymerase chain reaction qrt pcr validation
    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by <t>qRT-PCR.</t> (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr Validation, supplied by Genewiz, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcription polymerase chain reaction qrt pcr validation/product/Genewiz
    Average 95 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    reverse transcription polymerase chain reaction qrt pcr validation - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    86
    Stratagene qrt pcr analysis polymerase chain reactions
    MYC3 and MYC4 regulate expression of JA-responsive genes. (A, B) <t>qRT-PCR</t> expression analysis of JAZ genes and wound-responsive genes in 10-d-old wild-type and MYC overexpression seedlings without JA treatment. Wild-type samples served as a calibrator for the calculation of relative expression levels (arbitrarily set to one). (C) Relative transcript levels of the PDF1.2 gene in 10-d-old wild-type and overexpression transgenic seedlings with or without 10 μM JA treatment (incubated for 6 h). Wild-type samples without JA treatment served as a calibrator, and relative expression was determined from replicate measurements in two independent biological replicates. Data are mean ±SE.
    Qrt Pcr Analysis Polymerase Chain Reactions, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr analysis polymerase chain reactions/product/Stratagene
    Average 86 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis polymerase chain reactions - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    94
    TaKaRa transcription polymerase chain reaction qrt pcr
    MYC3 and MYC4 regulate expression of JA-responsive genes. (A, B) <t>qRT-PCR</t> expression analysis of JAZ genes and wound-responsive genes in 10-d-old wild-type and MYC overexpression seedlings without JA treatment. Wild-type samples served as a calibrator for the calculation of relative expression levels (arbitrarily set to one). (C) Relative transcript levels of the PDF1.2 gene in 10-d-old wild-type and overexpression transgenic seedlings with or without 10 μM JA treatment (incubated for 6 h). Wild-type samples without JA treatment served as a calibrator, and relative expression was determined from replicate measurements in two independent biological replicates. Data are mean ±SE.
    Transcription Polymerase Chain Reaction Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transcription polymerase chain reaction qrt pcr/product/TaKaRa
    Average 94 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    87
    TaKaRa polymerase chain reaction qrt pcr analysis
    MYC3 and MYC4 regulate expression of JA-responsive genes. (A, B) <t>qRT-PCR</t> expression analysis of JAZ genes and wound-responsive genes in 10-d-old wild-type and MYC overexpression seedlings without JA treatment. Wild-type samples served as a calibrator for the calculation of relative expression levels (arbitrarily set to one). (C) Relative transcript levels of the PDF1.2 gene in 10-d-old wild-type and overexpression transgenic seedlings with or without 10 μM JA treatment (incubated for 6 h). Wild-type samples without JA treatment served as a calibrator, and relative expression was determined from replicate measurements in two independent biological replicates. Data are mean ±SE.
    Polymerase Chain Reaction Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 87/100, based on 9 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 87 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    90
    Bio-Rad quantitative rt polymerase chain reaction qrt pcr reactions
    PUM1 regulates ATXN1 levels via a highly conserved binding motif (A) Schematic representation of Human ATXN1 -3′UTR showing three putative PUM1 binding motifs (gray and red boxes) and their conservation in different species. The numbers indicate positions of PUM1 motifs in the human ATXN1 -3′UTR. (B) ATXN1 mRNA quantification by <t>qRT-PCR</t> in HEK293T cells upon overexpression (left panel) or knockdown (si PUM1 -81 and -82) (right panel) of PUM1 . The destination-cloning vector (Control), scrambled siRNAs (siScr.) and cel-miR-67 were used as negative controls. The housekeeping gene GAPDH was used to normalize the expression of genes in all the qRT-PCR experiments. (C) Luciferase assay in HEK293 cells overexpressing the reporter construct harboring the full length ATXN1 -3′UTR. In these conditions, we overexpressed (left panel) or decreased expression (right panel) of PUM1 . (D) Luciferase assay in HEK293T cells transfected with single WT and mutant (MUT) putative PUM1 binding site on the ATXN1 -3′UTR. The position of cloned regions for each PUM1 binding motif are indicated. The mutagenized nucleotides are highlighted in blue. In panels C and D the destination-vector (control), RNAi scramble (siScramble) and cel-miR-67 were used as negative controls; miR-101a was used as positive control. (RL) Renilla, (FL) Firefly luciferase. All the experiments were performed in triplicate (data represent mean ± S.E.M.). P values were calculated by Student’s t-test. Statistical significance is indicated as follows: * P
    Quantitative Rt Polymerase Chain Reaction Qrt Pcr Reactions, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative rt polymerase chain reaction qrt pcr reactions/product/Bio-Rad
    Average 90 stars, based on 34 article reviews
    Price from $9.99 to $1999.99
    quantitative rt polymerase chain reaction qrt pcr reactions - by Bioz Stars, 2020-04
    90/100 stars
      Buy from Supplier

    99
    Thermo Fisher quantitative reverse transcription polymerase chain reaction qrt pcr
    ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by <t>qRT-PCR</t> (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.
    Quantitative Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1303 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative reverse transcription polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 99 stars, based on 1303 article reviews
    Price from $9.99 to $1999.99
    quantitative reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    89
    Roche transcription polymerase chain reaction qrt pcr
    ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by <t>qRT-PCR</t> (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.
    Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transcription polymerase chain reaction qrt pcr/product/Roche
    Average 89 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    89/100 stars
      Buy from Supplier

    91
    Thermo Fisher real time polymerase chain reaction qrt pcr amplification
    Characterization of apoptosis triggered by TCEA3 expression. a RH30 cells expressing TCEA3 or empty vector control were harvested for RNA. BAX and BCL2 gene expression were assayed by <t>qRT-PCR.</t> Scale bars 50 µm. b RH30 cells expressing TCEA3 or empty vector were assayed by western blot with antibodies against the indicated apoptotic marker proteins. GAPDH was used as a loading control. c RH30 cells expressing TCEA3 and empty vector were treated with caspase 8 inhibitor (Z-IETD-FMK, 40 μM), caspase 9 inhibitor (Z-LEHD-FMK, 40 μM) or a pan-caspase inhibitor (Z-VAD-FMK, 50 μM) for 18 h. Immunofluorescence assay was done with anti TCEA3 antibody (green) and anti-cleaved caspase 3 antibody (red). DAPI (blue) was used to visualize nuclei. Scale bar is 50 µm. d Western blot assay on the same cells as in C. to confirm the protein expression of TCEA3, caspase 3 and tubulin as a loading control. * marks cleaved caspase 3. Error bars are S.E.M. Student t test; *** p
    Real Time Polymerase Chain Reaction Qrt Pcr Amplification, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr amplification/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr amplification - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    88
    TaKaRa quantitative polymerase chain reaction qrt pcr reactions
    Pluripotency of in vivo rPSCs. ( A ) The established in vivo rPSCs expressed Oct4 -GFP and were morphologically indistinguishable from mESCs. Scale bar = 100 μm. ( B ) Immunocytochemistry analysis (scale bar = 50 μm) and ( C ) <t>qRT-PCR</t> showed that in vivo rPSCs expressed pluripotency markers Oct4 (endo), Nanog (endo), Sox2 , and Rex1 , similar to in vitro iPSCs and ESCs. ( D ) DNA methylation status of Oct4 and Nanog promoter region in rOG2-T-rPSCs #1, #2. ( E,F ) The rOG2-T-rPSCs could differentiate into all 3 germ layers in vitro and in vivo Scale bar = 100 μm. ( G ) Aggregation potential of in viv o rPSCs into the inner cell mass of normal embryos. The in vivo rPSCs did not contribute to the trophectoderm lineage. Scale bar = 50 μm.
    Quantitative Polymerase Chain Reaction Qrt Pcr Reactions, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative polymerase chain reaction qrt pcr reactions/product/TaKaRa
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    quantitative polymerase chain reaction qrt pcr reactions - by Bioz Stars, 2020-04
    88/100 stars
      Buy from Supplier

    99
    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 99 stars, based on 2185 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    94
    Thermo Fisher quantificational reverse transcription polymerase chain reaction qrt pcr trizol
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Quantificational Reverse Transcription Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantificational reverse transcription polymerase chain reaction qrt pcr trizol/product/Thermo Fisher
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    quantificational reverse transcription polymerase chain reaction qrt pcr trizol - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    94
    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr trizol
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, 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/quantitative real time polymerase chain reaction qrt pcr trizol/product/Thermo Fisher
    Average 94 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr trizol - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    86
    Solexa real time polymerase chain reaction quantitative real time polymerase chain reaction qrt pcr
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Real Time Polymerase Chain Reaction Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Solexa, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction quantitative real time polymerase chain reaction qrt pcr/product/Solexa
    Average 86 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    86
    Bio-Rad quantitative reverse polymerase chain reaction qrt pcr
    Genome-wide analysis for cryptic SVs identifies disruption of further neurodevelopmental disease candidate genes and demonstrates reduced expression of ZNF804A in patient cells. (A,B) Graphical representation of the three genes disrupted by an ∼10 kB deletion on chr 19 [del(19q13.4)] (A) and the cryptic paracentric inversion inv(2)(p32.1q32.1) in the patient. Sites of breakpoints are denoted by arrows. (C) Graphical representation of anomalous-read (red dots) fusion positions for the cryptic 2.49 Mb paracentric inversion on chromosome 2. Mate-pair library sequencing-predicted breakpoints 5′ and 3′ of the inversion were amplified with breakpoint-specific primers and validated at base-pair level by <t>PCR</t> and capillary sequencing. (D) mRNA-levels of ZNF804A and the housekeeping gene RPL19 were quantified by <t>qRT-PCR</t> from total RNA isolated from fibroblasts of the patient or a healthy male control and normalized to expression of beta-actin.
    Quantitative Reverse Polymerase Chain Reaction Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative reverse polymerase chain reaction qrt pcr/product/Bio-Rad
    Average 86 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    quantitative reverse polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    95
    Roche reverse transcription polymerase chain reaction qrt pcr reaction
    Analysis of HIC1 expression in colorectal neoplasia. (A) HIC1 expression changes during tumor progression. <t>qRT-PCR</t> analysis of the HIC1 mRNA levels in healthy tissue, hyperplastic adenomas (Hyp; n = 9), adenomas displaying low-grade (LGD; n = 24) or high-grade (HGD; n = 25) dysplasia, and CRC ( n = 12). The boxed areas correspond to the second and third quartiles; the median of ΔCp values for each category is indicated as the red line. The relation between the HIC1 expression profile and neoplasia progression is significant, as evidenced by the Spearman’s ( ρ = − 0.67) and Kendall’s ( τ = − 0.51) coefficient values. (B) HIC1 expression in CRC subgroups clustered according to the DNA methylation profiles. HIC1-high and HIC1-low samples are indicated by brown or blue dots, respectively. Black dots indicate the other specimens. The red lines correspond to the median values; log2-expression intensity, binary logarithm of expression intensity (additional details are given in Supplementary Methods).
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr Reaction, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcription polymerase chain reaction qrt pcr reaction/product/Roche
    Average 95 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    reverse transcription polymerase chain reaction qrt pcr reaction - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    92
    TaKaRa quantitative reverse transcriptase polymerase chain reaction qrt pcr kit
    Analysis of HIC1 expression in colorectal neoplasia. (A) HIC1 expression changes during tumor progression. <t>qRT-PCR</t> analysis of the HIC1 mRNA levels in healthy tissue, hyperplastic adenomas (Hyp; n = 9), adenomas displaying low-grade (LGD; n = 24) or high-grade (HGD; n = 25) dysplasia, and CRC ( n = 12). The boxed areas correspond to the second and third quartiles; the median of ΔCp values for each category is indicated as the red line. The relation between the HIC1 expression profile and neoplasia progression is significant, as evidenced by the Spearman’s ( ρ = − 0.67) and Kendall’s ( τ = − 0.51) coefficient values. (B) HIC1 expression in CRC subgroups clustered according to the DNA methylation profiles. HIC1-high and HIC1-low samples are indicated by brown or blue dots, respectively. Black dots indicate the other specimens. The red lines correspond to the median values; log2-expression intensity, binary logarithm of expression intensity (additional details are given in Supplementary Methods).
    Quantitative Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative reverse transcriptase polymerase chain reaction qrt pcr kit/product/TaKaRa
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    quantitative reverse transcriptase polymerase chain reaction qrt pcr kit - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    Image Search Results


    HIF-1α regulated the expression of CCL2 in PDAC cells, PC, N and H indicate positive control, normoxia, and hypoxia, respectively. ( A ) HPDE6-C7,BxPC3 and MiaPaca-2 cells were transfected with siHIF-1α (50 nmol/L) and pcDNA3.1-HIF-1α plasmids (4 µg) for 48 h, respectively; the mRNA expression levels of HIF-1α and CCL2 were assessed by qRT-PCR. The experiments were performed thrice independently. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Hypoxia Inducible Factor 1 (HIF-1) Recruits Macrophage to Activate Pancreatic Stellate Cells in Pancreatic Ductal Adenocarcinoma

    doi: 10.3390/ijms17060799

    Figure Lengend Snippet: HIF-1α regulated the expression of CCL2 in PDAC cells, PC, N and H indicate positive control, normoxia, and hypoxia, respectively. ( A ) HPDE6-C7,BxPC3 and MiaPaca-2 cells were transfected with siHIF-1α (50 nmol/L) and pcDNA3.1-HIF-1α plasmids (4 µg) for 48 h, respectively; the mRNA expression levels of HIF-1α and CCL2 were assessed by qRT-PCR. The experiments were performed thrice independently. * p

    Article Snippet: The reverse transcription polymerase chain reaction (qRT-PCR) system (TaKaRa Bio Group, Shiga, Japan) was utilized to obtain complementary DNA (cDNA).

    Techniques: Expressing, Positive Control, Transfection, Quantitative RT-PCR

    Primer Design and qRT-PCR

    Journal: Plastic and reconstructive surgery

    Article Title: Beta-catenin-dependent Wnt signaling: a pathway in acute cutaneous wounding

    doi: 10.1097/PRS.0000000000004170

    Figure Lengend Snippet: Primer Design and qRT-PCR

    Article Snippet: All other reagents for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Applied Biosystems (Foster City, CA). rmWnt3a was from R & D Systems (Minneapolis, MN).

    Techniques: Quantitative RT-PCR

    The effect of TGF-β3 by TCDD induced in MEPM cells. 0.5 nM, 1 nM, 5 nM, 10 nM, 20 nM and 50 nM TCDD, or DMSO (≤0.05%)-treated MEPM cells as the experiment group and control group, respectively. After treated for 72 hours, the expression of TGF-β3 was measured by qRT-PCR. Data were mean values ± standard deviation of 3 replicate experiments. * P

    Journal: Dose-Response

    Article Title: 2,3,7,8-Tetrachlorodibenzo-p-Dioxin and TGF-β3 Mediated-Mouse Embryonic Palatal Mesenchymal Cells

    doi: 10.1177/1559325818810637

    Figure Lengend Snippet: The effect of TGF-β3 by TCDD induced in MEPM cells. 0.5 nM, 1 nM, 5 nM, 10 nM, 20 nM and 50 nM TCDD, or DMSO (≤0.05%)-treated MEPM cells as the experiment group and control group, respectively. After treated for 72 hours, the expression of TGF-β3 was measured by qRT-PCR. Data were mean values ± standard deviation of 3 replicate experiments. * P

    Article Snippet: To detect expression of TGF-β3, first strand complementary DNA (cDNA) was synthesized using a PrimeScript II 1st strand cDNA Synthesis Kit (6210A, TakaRa Biotechnology, Kyoto, Japan), and then amplified by quantitative real-time polymerase chain reaction (qRT-PCR) with the SYBR Premix Ex Taq Kit (DRR420A; TaKaRa) through ABI 7900 PRISM system (7900HT; Applied Biosystems, Carlsbad, California).

    Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

    CDKI expression in human terminal erythroid differentiation. (A) RNA-seq data showing CDKI members’ expression (fragments per kilobase of transcript per million) at each distinct stage of human terminal erythroid differentiation. Baso-E, basophilic erythroblast; ortho-E, orthochromatic erythroblast; poly-E, polychromatic erythroblast; pro-E, proerythroblast. (B) Representative image of western blotting of p19 INK4d , p18 INK4c , and p27 KIP1 expression in whole-cell lysates prepared from erythroblasts cultured for different times (left). Quantitative analysis of data from 3 independent experiments of protein expression levels (right). GAPDH was used as a loading control. (C) qRT-PCR results showing p19 INK4d expression in erythroblasts infected with lentivirus containing control (Lucif shRNA) or p19 INK4d shRNA on day (D) 7, D 11, and D 15 of culture. The results were normalized to GAPDH mRNA. (D) Representative images of western blotting showing p19 INK4d expression levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (left). Quantitative analysis of protein expression data from 3 independent experiments (right). The results were normalized to GAPDH protein. Statistical analysis of data of 3 independent experiments and bar plot represents mean ± SD of triplicate samples. *** P

    Journal: Blood

    Article Title: Unexpected role for p19INK4d in posttranscriptional regulation of GATA1 and modulation of human terminal erythropoiesis

    doi: 10.1182/blood-2016-09-739268

    Figure Lengend Snippet: CDKI expression in human terminal erythroid differentiation. (A) RNA-seq data showing CDKI members’ expression (fragments per kilobase of transcript per million) at each distinct stage of human terminal erythroid differentiation. Baso-E, basophilic erythroblast; ortho-E, orthochromatic erythroblast; poly-E, polychromatic erythroblast; pro-E, proerythroblast. (B) Representative image of western blotting of p19 INK4d , p18 INK4c , and p27 KIP1 expression in whole-cell lysates prepared from erythroblasts cultured for different times (left). Quantitative analysis of data from 3 independent experiments of protein expression levels (right). GAPDH was used as a loading control. (C) qRT-PCR results showing p19 INK4d expression in erythroblasts infected with lentivirus containing control (Lucif shRNA) or p19 INK4d shRNA on day (D) 7, D 11, and D 15 of culture. The results were normalized to GAPDH mRNA. (D) Representative images of western blotting showing p19 INK4d expression levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (left). Quantitative analysis of protein expression data from 3 independent experiments (right). The results were normalized to GAPDH protein. Statistical analysis of data of 3 independent experiments and bar plot represents mean ± SD of triplicate samples. *** P

    Article Snippet: RNA extracted from primary cultured human erythroid cells using TRIzol reagent (Invitrogen) was reverse transcripted with SuperScript first-strand synthesis system (Invitrogen) and amplified by quantitative polymerase chain reaction (qRT-PCR) with the Cycler (Bio-Rad) and appropriate primer pairs.

    Techniques: Expressing, RNA Sequencing Assay, Western Blot, Cell Culture, Quantitative RT-PCR, Infection, shRNA

    p19 INK4d knockdown decreased GATA1 protein expression and ectopic GATA1 expression rescues the differentiation delay. (A) Representative images of western blotting analysis showing GATA1, 4.1R, tropomodulin (Tmod), HBG, p18 INK4c , KLF1, and CD44 expression in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (day 11 cells) (left). Quantitative analysis of protein expression data from 3 independent experiments (right). GAPDH was used as a loading control and the results were normalized to GAPDH protein expression. (B) qRT-PCR analysis of GATA1 mRNA levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA. The results were normalized to GAPDH mRNA. (C) Representative data of flow cytometry analysis of erythroblasts infected with Lucif shRNA, p19 INK4d shRNA, and p19 INK4d shRNA combined with GATA1 overexpression. GPA expression was monitored on D 9 cells (left) and expression of band 3 and α4-integrin on D 15 cells (right). The red numbers indicate statistical analysis of GPA-positive rate from 3 independent experiments. (D) Representative images of western blotting showing GATA1 levels in whole cell lysates prepared from erythroblasts (left). Quantitative analysis of expression data from 3 independent experiments (right). GAPDH was used as a loading control. Statistical analysis of data from 3 independent experiments and bar plot represents mean ± SD of triplicate samples. * P

    Journal: Blood

    Article Title: Unexpected role for p19INK4d in posttranscriptional regulation of GATA1 and modulation of human terminal erythropoiesis

    doi: 10.1182/blood-2016-09-739268

    Figure Lengend Snippet: p19 INK4d knockdown decreased GATA1 protein expression and ectopic GATA1 expression rescues the differentiation delay. (A) Representative images of western blotting analysis showing GATA1, 4.1R, tropomodulin (Tmod), HBG, p18 INK4c , KLF1, and CD44 expression in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA (day 11 cells) (left). Quantitative analysis of protein expression data from 3 independent experiments (right). GAPDH was used as a loading control and the results were normalized to GAPDH protein expression. (B) qRT-PCR analysis of GATA1 mRNA levels in erythroblasts infected with Lucif shRNA or p19 INK4d shRNA. The results were normalized to GAPDH mRNA. (C) Representative data of flow cytometry analysis of erythroblasts infected with Lucif shRNA, p19 INK4d shRNA, and p19 INK4d shRNA combined with GATA1 overexpression. GPA expression was monitored on D 9 cells (left) and expression of band 3 and α4-integrin on D 15 cells (right). The red numbers indicate statistical analysis of GPA-positive rate from 3 independent experiments. (D) Representative images of western blotting showing GATA1 levels in whole cell lysates prepared from erythroblasts (left). Quantitative analysis of expression data from 3 independent experiments (right). GAPDH was used as a loading control. Statistical analysis of data from 3 independent experiments and bar plot represents mean ± SD of triplicate samples. * P

    Article Snippet: RNA extracted from primary cultured human erythroid cells using TRIzol reagent (Invitrogen) was reverse transcripted with SuperScript first-strand synthesis system (Invitrogen) and amplified by quantitative polymerase chain reaction (qRT-PCR) with the Cycler (Bio-Rad) and appropriate primer pairs.

    Techniques: Expressing, Western Blot, Infection, shRNA, Quantitative RT-PCR, Flow Cytometry, Cytometry, Over Expression

    LPS / ATP activate NLRP 3 inflammasomes in VSMC s. Human primary VSMC s were treated with vehicle or the indicated concentrations of LPS for 16 hours and/or ATP for 1 hour. Some cells were transfected with control or NLRP 3‐specific si RNA or treated with vehicle DMSO or ZYVAD ‐ FMK , followed by treatment with LPS and/or ATP . The relative levels of NLRP 3 and cleaved caspase‐1 expression were determined by qRT ‐ PCR and Western blot. The distribution of HMGB 1 in the different groups of cells and their cultured supernatants were determined by Western blot and ELISA . Data are representative images or expressed as the mean± SD of each group from 4 separated experiments (Western blot) or 3 separated experiments with 4 duplicated wells ( qRT ‐ PCR and ELISA ). A, The levels of NLRP 3 mRNA transcripts. B, The levels of NLRP 3 proteins. C, The levels of cleaved caspase‐1. D, The levels of total HMGB 1. E, The levels of nuclear HMGB 1. F, The levels of cytoplasmic HMGB 1. G, The levels of HMGB 1 in the supernatants. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Activation of NLRP3 Inflammasome Promotes Foam Cell Formation in Vascular Smooth Muscle Cells and Atherogenesis Via HMGB1

    doi: 10.1161/JAHA.118.008596

    Figure Lengend Snippet: LPS / ATP activate NLRP 3 inflammasomes in VSMC s. Human primary VSMC s were treated with vehicle or the indicated concentrations of LPS for 16 hours and/or ATP for 1 hour. Some cells were transfected with control or NLRP 3‐specific si RNA or treated with vehicle DMSO or ZYVAD ‐ FMK , followed by treatment with LPS and/or ATP . The relative levels of NLRP 3 and cleaved caspase‐1 expression were determined by qRT ‐ PCR and Western blot. The distribution of HMGB 1 in the different groups of cells and their cultured supernatants were determined by Western blot and ELISA . Data are representative images or expressed as the mean± SD of each group from 4 separated experiments (Western blot) or 3 separated experiments with 4 duplicated wells ( qRT ‐ PCR and ELISA ). A, The levels of NLRP 3 mRNA transcripts. B, The levels of NLRP 3 proteins. C, The levels of cleaved caspase‐1. D, The levels of total HMGB 1. E, The levels of nuclear HMGB 1. F, The levels of cytoplasmic HMGB 1. G, The levels of HMGB 1 in the supernatants. * P

    Article Snippet: The relative levels of target gene mRNA transcripts were determined by quantitative real‐time polymerase chain reaction (qRT‐PCR) using the FastStart Universal SYBR Green Master (Roche Applied Sciences) and specific primers on a Roche LightCycler 480 Real Time PCR System.

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

    NLRP 3 inflammasome activation reduces LXR α and ABCA 1 expression in VSMC s. Following treatment with the indicated drugs and LPS / ATP stimulation, the relative levels of LXR α and ABCA 1 expression were determined qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR ). A, qRT ‐ PCR analysis of LXR α and ABCA 1 mRNA transcripts. B, Western blot analysis of LXR α and ABCA 1 proteins. C through F, Treatment with ZYVAD ‐ FMK or NLRP 3 silencing mitigated the LPS / ATP ‐reduced LXR α and ABCA 1 expression. G and H, Treatment with T091317, the LXR α agonist enhanced ABCA 1 expression and mitigated the LPS / ATP ‐reduced ABCA 1 expression. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Activation of NLRP3 Inflammasome Promotes Foam Cell Formation in Vascular Smooth Muscle Cells and Atherogenesis Via HMGB1

    doi: 10.1161/JAHA.118.008596

    Figure Lengend Snippet: NLRP 3 inflammasome activation reduces LXR α and ABCA 1 expression in VSMC s. Following treatment with the indicated drugs and LPS / ATP stimulation, the relative levels of LXR α and ABCA 1 expression were determined qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR ). A, qRT ‐ PCR analysis of LXR α and ABCA 1 mRNA transcripts. B, Western blot analysis of LXR α and ABCA 1 proteins. C through F, Treatment with ZYVAD ‐ FMK or NLRP 3 silencing mitigated the LPS / ATP ‐reduced LXR α and ABCA 1 expression. G and H, Treatment with T091317, the LXR α agonist enhanced ABCA 1 expression and mitigated the LPS / ATP ‐reduced ABCA 1 expression. * P

    Article Snippet: The relative levels of target gene mRNA transcripts were determined by quantitative real‐time polymerase chain reaction (qRT‐PCR) using the FastStart Universal SYBR Green Master (Roche Applied Sciences) and specific primers on a Roche LightCycler 480 Real Time PCR System.

    Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Western Blot

    HMGB 1 reduces LXR α and ABCA 1 expression in VSMC s. Following pretreatment with HMGB 1 and/or glycyrrhizin the HMGB 1 inhibitor and LPS / ATP stimulation, the relative levels of LXR α and ABCA 1 expression were determined by qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR ). A and B, inhibition of HMGB 1 mitigated the LPS / ATP ‐reduced LXR α and ABCA 1 expression. C and D, HMGB 1 reduces the levels of LXR α and ABCA 1 expression in a dose‐dependent manner. E and F, Treatment with T091317 mitigated the HMGB 1‐reduced ABCA 1 expression. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Activation of NLRP3 Inflammasome Promotes Foam Cell Formation in Vascular Smooth Muscle Cells and Atherogenesis Via HMGB1

    doi: 10.1161/JAHA.118.008596

    Figure Lengend Snippet: HMGB 1 reduces LXR α and ABCA 1 expression in VSMC s. Following pretreatment with HMGB 1 and/or glycyrrhizin the HMGB 1 inhibitor and LPS / ATP stimulation, the relative levels of LXR α and ABCA 1 expression were determined by qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR ). A and B, inhibition of HMGB 1 mitigated the LPS / ATP ‐reduced LXR α and ABCA 1 expression. C and D, HMGB 1 reduces the levels of LXR α and ABCA 1 expression in a dose‐dependent manner. E and F, Treatment with T091317 mitigated the HMGB 1‐reduced ABCA 1 expression. * P

    Article Snippet: The relative levels of target gene mRNA transcripts were determined by quantitative real‐time polymerase chain reaction (qRT‐PCR) using the FastStart Universal SYBR Green Master (Roche Applied Sciences) and specific primers on a Roche LightCycler 480 Real Time PCR System.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Inhibition

    NLRP 3 inflammasome‐dependent HMGB 1 secretion reduces LXR α and ABCA 1 expression through binding to RAGE in VSMC s. Following NLRP 3 silencing, pretreatment with RAP , a RAGE antagonist peptide and LPS / ATP stimulation in the presence or absence of Chol:Mβ CD , the cholesterol accumulation in VSMC s was characterized and the relative levels of LXR α and ABCA 1 expression were determined by qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR and Cholesterol Efflux Fluorometric Assay). A through C, The cholesterol accumulation in VSMC s. D, The cholesterol efflux in VSMC s. E through H, The relative levels of LXR α and ABCA 1 expression in the different groups of VSMC s. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Activation of NLRP3 Inflammasome Promotes Foam Cell Formation in Vascular Smooth Muscle Cells and Atherogenesis Via HMGB1

    doi: 10.1161/JAHA.118.008596

    Figure Lengend Snippet: NLRP 3 inflammasome‐dependent HMGB 1 secretion reduces LXR α and ABCA 1 expression through binding to RAGE in VSMC s. Following NLRP 3 silencing, pretreatment with RAP , a RAGE antagonist peptide and LPS / ATP stimulation in the presence or absence of Chol:Mβ CD , the cholesterol accumulation in VSMC s was characterized and the relative levels of LXR α and ABCA 1 expression were determined by qRT ‐ PCR and Western blot. Data are representative images or expressed as the mean± SD of each group from 3 separated experiments (Western blot) or 3 separated experiments with 3 duplicated wells ( qRT ‐ PCR and Cholesterol Efflux Fluorometric Assay). A through C, The cholesterol accumulation in VSMC s. D, The cholesterol efflux in VSMC s. E through H, The relative levels of LXR α and ABCA 1 expression in the different groups of VSMC s. * P

    Article Snippet: The relative levels of target gene mRNA transcripts were determined by quantitative real‐time polymerase chain reaction (qRT‐PCR) using the FastStart Universal SYBR Green Master (Roche Applied Sciences) and specific primers on a Roche LightCycler 480 Real Time PCR System.

    Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Western Blot

    Elevated levels of Mg 2+ in APP/PS1 transgenic mice decrease the expression of IL-1β. The APP/PS1 transgenic mice at the age of 4 months were administered Mg 2+ (100 mg/kg/d) for 2 months before collecting the brain ( a ). In select experiments, the brains of APP/PS1 transgenic mice at the age of 3 months were harvested and sectioned (400 μm) using a cryostat ( b ). In separate experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) and the right cerebral ventricle was injected (i.c.v.) with D1A cells, which was pre-transfected with IL-1β promoter in the right cerebral ventricle ( c ). In distinct experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) before staining with IL-1β antibody and scanning under two-photon microscopy ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody (a left panel, b). These images are representative of six independent experiments, all with similar results. IL-1β protein and mRNA levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively (a right panel). The total amounts of GAPDH and protein served as an internal control. The experimental cartoon and real surgery images are shown (c, d upper panel). Luciferase activities from the different groups of mice were measured using a live animal imaging system (c lower panel). The immunofluorescence of IL-1β was scanned using a two-photon microscope (d lower panel). The data represent the means ± S.E. of three independent experiments. * p

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: Elevated levels of Mg 2+ in APP/PS1 transgenic mice decrease the expression of IL-1β. The APP/PS1 transgenic mice at the age of 4 months were administered Mg 2+ (100 mg/kg/d) for 2 months before collecting the brain ( a ). In select experiments, the brains of APP/PS1 transgenic mice at the age of 3 months were harvested and sectioned (400 μm) using a cryostat ( b ). In separate experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) and the right cerebral ventricle was injected (i.c.v.) with D1A cells, which was pre-transfected with IL-1β promoter in the right cerebral ventricle ( c ). In distinct experiments, the left cerebral ventricle was injected with Mg 2+ (2 μg/5 μl) or vehicle (PBS) before staining with IL-1β antibody and scanning under two-photon microscopy ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody (a left panel, b). These images are representative of six independent experiments, all with similar results. IL-1β protein and mRNA levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively (a right panel). The total amounts of GAPDH and protein served as an internal control. The experimental cartoon and real surgery images are shown (c, d upper panel). Luciferase activities from the different groups of mice were measured using a live animal imaging system (c lower panel). The immunofluorescence of IL-1β was scanned using a two-photon microscope (d lower panel). The data represent the means ± S.E. of three independent experiments. * p

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

    Techniques: Transgenic Assay, Mouse Assay, Expressing, Injection, Transfection, Staining, Microscopy, Immunohistochemistry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Luciferase, Imaging, Immunofluorescence

    MgT treatment attenuated the effects of Aβ oligomers in the CSF for inducing the expression of IL-1β in the cerebral cortex. The WT C57BL/6 mice at the age of 3 months were injected (i.c.v) with Aβ oligomers (2 μg/5 μl) in the absence or presence of Mg 2+ (2 μg/5 μl). The brains were then collected and sectioned after 24 h ( a and b ). In select experiments, the brains of WT C57BL/6 mice at the age of 3 months were harvested and freshly sectioned (400 μm) before treatment with Aβ oligomers (1 μM) in the absence or presence of MgT (50 μM) for 24 h ( c ). In distinct experiments, A172 cells were treated with Aβ oligomers (1 μM) in the absence or presence of MgT (50 μM) for 24 h ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody ( a and c ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively (b and d). The total amounts of GAPDH and protein served as an internal control. The data represent the means ± SE of three independent experiments. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: MgT treatment attenuated the effects of Aβ oligomers in the CSF for inducing the expression of IL-1β in the cerebral cortex. The WT C57BL/6 mice at the age of 3 months were injected (i.c.v) with Aβ oligomers (2 μg/5 μl) in the absence or presence of Mg 2+ (2 μg/5 μl). The brains were then collected and sectioned after 24 h ( a and b ). In select experiments, the brains of WT C57BL/6 mice at the age of 3 months were harvested and freshly sectioned (400 μm) before treatment with Aβ oligomers (1 μM) in the absence or presence of MgT (50 μM) for 24 h ( c ). In distinct experiments, A172 cells were treated with Aβ oligomers (1 μM) in the absence or presence of MgT (50 μM) for 24 h ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody ( a and c ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively (b and d). The total amounts of GAPDH and protein served as an internal control. The data represent the means ± SE of three independent experiments. * P

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

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

    Involvement of ERK1/2 and PPARγ pathways in regulating the expression of IL-1β in MgT-treated A172 or D1A cells. Human glioblastoma A172 ( a ) or mouse astrocytes/microglia D1A cells ( b ) were treated with MgT (50 μM) for 48 h. In select experiments, A172 cells were treated with PD98059 (10 μM) in the absence or presence of MgT (50 μM) for 48 h ( c and e ). In separate experiments, the cells were transfected with ERK1/2 ( d ) or PPARγ siRNA ( f ) before incubation with MgT (50 μM) for 48 h. In distinct experiments, A172 cells were treated with GW9662 (1 μM) in the absence or presence of MgT (50 μM) for 48 h ( e '). In other experiments, primary cultured astrocytes were treated with MgT (50 μM) in the absence or presence of PD98059 (10 μM) (g) or GW9662 (1 μM) for 48 h (h). Total ERK1/2 ( c , d and g upper panel), phosphorylated ERK1/2 levels ( c and g upper panel), total PPARγ ( e and f upper panel), and phosphorylated PPARγ ( e ) were detected by immunoblotting using specific Abs. Equal lane loading is demonstrated by the similar intensities of total β-actin. IL-1β protein and mRNA levels were determined by IL-1β enzyme immunoassay kits and qRT-PCR, respectively. The total amounts of protein and GAPDH served as internal controls. The data represent the means ± SE of three independent experiments. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: Involvement of ERK1/2 and PPARγ pathways in regulating the expression of IL-1β in MgT-treated A172 or D1A cells. Human glioblastoma A172 ( a ) or mouse astrocytes/microglia D1A cells ( b ) were treated with MgT (50 μM) for 48 h. In select experiments, A172 cells were treated with PD98059 (10 μM) in the absence or presence of MgT (50 μM) for 48 h ( c and e ). In separate experiments, the cells were transfected with ERK1/2 ( d ) or PPARγ siRNA ( f ) before incubation with MgT (50 μM) for 48 h. In distinct experiments, A172 cells were treated with GW9662 (1 μM) in the absence or presence of MgT (50 μM) for 48 h ( e '). In other experiments, primary cultured astrocytes were treated with MgT (50 μM) in the absence or presence of PD98059 (10 μM) (g) or GW9662 (1 μM) for 48 h (h). Total ERK1/2 ( c , d and g upper panel), phosphorylated ERK1/2 levels ( c and g upper panel), total PPARγ ( e and f upper panel), and phosphorylated PPARγ ( e ) were detected by immunoblotting using specific Abs. Equal lane loading is demonstrated by the similar intensities of total β-actin. IL-1β protein and mRNA levels were determined by IL-1β enzyme immunoassay kits and qRT-PCR, respectively. The total amounts of protein and GAPDH served as internal controls. The data represent the means ± SE of three independent experiments. * P

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

    Techniques: Expressing, Transfection, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    Aβ oligomers in the CSF of APP/PS1 mice have the ability to stimulate the expression of IL-1β in the cerebral cortex of WT mice. The APP/PS1 Tg mice at the age of 4 months were treated with Mg 2+ (100 mg/kg/d) for 5 months before collecting the CSF and brains ( a-c ). Cerebrospinal fluid (CSF) was then injected into the wild type C57BL/6 mice in the absence or presence of Aβ antibody (1 μg/5 μl) for two weeks before scarifice ( d-f ). The production of Aβ 1-42 was determined by ELISA kits, and the total amount of protein served as an internal control ( a ). The immunoreactivity of Aβ and IL-1β was determined by immunohistochemistry using an anti-Aβ or IL-1β antibody ( c, d ). These images are representative of six independent experiments, all with similar results. The number of AP/fields was calculated according to the images of IHC ( b ). IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively ( e, f ). The total amounts of GAPDH and protein served as an internal control. * p

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: Aβ oligomers in the CSF of APP/PS1 mice have the ability to stimulate the expression of IL-1β in the cerebral cortex of WT mice. The APP/PS1 Tg mice at the age of 4 months were treated with Mg 2+ (100 mg/kg/d) for 5 months before collecting the CSF and brains ( a-c ). Cerebrospinal fluid (CSF) was then injected into the wild type C57BL/6 mice in the absence or presence of Aβ antibody (1 μg/5 μl) for two weeks before scarifice ( d-f ). The production of Aβ 1-42 was determined by ELISA kits, and the total amount of protein served as an internal control ( a ). The immunoreactivity of Aβ and IL-1β was determined by immunohistochemistry using an anti-Aβ or IL-1β antibody ( c, d ). These images are representative of six independent experiments, all with similar results. The number of AP/fields was calculated according to the images of IHC ( b ). IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively ( e, f ). The total amounts of GAPDH and protein served as an internal control. * p

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

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

    IL-1β was upregulated in AD patients and APP/PS1 transgenic mice. The tissue blocks of human brains at different stages of AD were collected from the New York Brain Bank at Columbia University. From 40 μm free-floating slices were prepared using a cryostat. ( a ) In select experiments, the brains of 3-month-old APP/PS1 transgenic mice were collected after anesthesia and perfusion. ( b – d ) The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody ( a and b ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA levels were determined by qRT-PCR, and total amounts of GAPDH served as an internal control ( c and d upper panels). The production of IL-1β in culture medium was determined using IL-1β enzyme immunoassay kits ( c and d lower panels). The data represent the means ± SE of three independent experiments. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: IL-1β was upregulated in AD patients and APP/PS1 transgenic mice. The tissue blocks of human brains at different stages of AD were collected from the New York Brain Bank at Columbia University. From 40 μm free-floating slices were prepared using a cryostat. ( a ) In select experiments, the brains of 3-month-old APP/PS1 transgenic mice were collected after anesthesia and perfusion. ( b – d ) The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β antibody ( a and b ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA levels were determined by qRT-PCR, and total amounts of GAPDH served as an internal control ( c and d upper panels). The production of IL-1β in culture medium was determined using IL-1β enzyme immunoassay kits ( c and d lower panels). The data represent the means ± SE of three independent experiments. * P

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

    Techniques: Transgenic Assay, Mouse Assay, Immunohistochemistry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    MgT treatment diminished the effects of IL-1β in the CSF with respect to inducing the expression of IL-1β in the cerebral cortex. WT C57BL/6 mice at the age of 3 months were intracerebroventricularly injected with IL-1β (0.5 μg/5 μl) in the absence or presence of Mg 2+ (2 μg/5 μl). The brains were then collected and sectioned after 24 h ( a and b ). In select experiments, the brains of WT C57BL/6 mice at the age of 3 months were harvested and freshly sectioned (400 μm) before treatment with IL-1β (100 ng ml –1 ) in the absence or presence of MgT (50 μM) for 24 h ( c ). In separate experiments, A172 cells were treated with IL-1β (100 ng ml –1 ) in the absence or presence of MgT (50 μM) for 24 h ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β or -APH-1 antibody ( a and c ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively ( b and d ). The total amounts of GAPDH and protein served as an internal control. The data represent the means ± SE of three independent experiments. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

    doi: 10.1038/cmi.2015.93

    Figure Lengend Snippet: MgT treatment diminished the effects of IL-1β in the CSF with respect to inducing the expression of IL-1β in the cerebral cortex. WT C57BL/6 mice at the age of 3 months were intracerebroventricularly injected with IL-1β (0.5 μg/5 μl) in the absence or presence of Mg 2+ (2 μg/5 μl). The brains were then collected and sectioned after 24 h ( a and b ). In select experiments, the brains of WT C57BL/6 mice at the age of 3 months were harvested and freshly sectioned (400 μm) before treatment with IL-1β (100 ng ml –1 ) in the absence or presence of MgT (50 μM) for 24 h ( c ). In separate experiments, A172 cells were treated with IL-1β (100 ng ml –1 ) in the absence or presence of MgT (50 μM) for 24 h ( d ). The immunoreactivity of IL-1β was determined by immunohistochemistry using an anti-IL-1β or -APH-1 antibody ( a and c ). These images are representative of six independent experiments, all with similar results. IL-1β mRNA and protein levels were determined by qRT-PCR and IL-1β enzyme immunoassay kits, respectively ( b and d ). The total amounts of GAPDH and protein served as an internal control. The data represent the means ± SE of three independent experiments. * P

    Article Snippet: All reagents for the quantitative real time polymerase chain reaction (qRT-PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments were purchased from Bio-Rad Laboratories (California, USA).

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

    The preventive effects of AMPK on the senescence associated with NAD + synthesis. (A) Cellular concentrations of NAD + on day 3 or day 5 after H 2 O 2 treatment. (B) Representative images of SA ‐β‐Gal staining of cells (left) and percentage of SA ‐β‐Gal‐positive cells (right). (C) Cellular concentrations of NAD + in the cells incubated with nicotinamide mononucleotide ( NMN ) for 3 days after H 2 O 2 treatment. (D) H 2 O 2 ‐treated cells were incubated with Met (10 mM ), BBR (10 μM), or CC (10 μM) for 3 days. The concentrations of NAD + are shown. (E) A schematic diagram of two pathways of NAD + synthesis. Red ones indicate rate‐limiting enzymes, and blue ones illustrate the enzyme inhibitors we used. (F) Relative fold‐changes in the mRNA levels of NAMPT , as monitored by qRT ‐ PCR (left), and representative images from immunoblot assays against NAMPT are shown (right). The ratio of NAMPT to actin was quantified by densitometry, based on the immunoblot images from three independent experiments. (G) Cells were incubated with Met, BBR , and CC for 3 days. Representative images from immunoblots against NAMPT are shown. The ratio of NAMPT to β‐actin was quantified by densitometry based on the immunoblot images from three independent experiments. (H) H 2 O 2 ‐treated NIH 3T3 cells were incubated with NAMPT inhibitor ( FK 866, 5 nM ), QPRT inhibitor ( PHTH , 1 mM ), Met (10 mM ), and BBR (10 μM) alone or in combination as indicated for 3 days. The percentages of SA ‐β‐Gal‐positive cells are shown. (I) The mRNA levels of NMNAT 1 , NMNAT 2 , and NMNAT 3 genes in the cells infected with corresponding lentivirus‐sh RNA or nontargeting sh RNA (sh CON ) as determined by qRT ‐ PCR . (J) Cellular concentrations of NAD + in sh RNA ‐infected cells. (K) Cells infected with sh RNA s were treated with H 2 O 2 and incubated with BBR for 3 days. Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). * P

    Journal: Aging Cell

    Article Title: AMPK activation protects cells from oxidative stress‐induced senescence via autophagic flux restoration and intracellular NAD+ elevation

    doi: 10.1111/acel.12446

    Figure Lengend Snippet: The preventive effects of AMPK on the senescence associated with NAD + synthesis. (A) Cellular concentrations of NAD + on day 3 or day 5 after H 2 O 2 treatment. (B) Representative images of SA ‐β‐Gal staining of cells (left) and percentage of SA ‐β‐Gal‐positive cells (right). (C) Cellular concentrations of NAD + in the cells incubated with nicotinamide mononucleotide ( NMN ) for 3 days after H 2 O 2 treatment. (D) H 2 O 2 ‐treated cells were incubated with Met (10 mM ), BBR (10 μM), or CC (10 μM) for 3 days. The concentrations of NAD + are shown. (E) A schematic diagram of two pathways of NAD + synthesis. Red ones indicate rate‐limiting enzymes, and blue ones illustrate the enzyme inhibitors we used. (F) Relative fold‐changes in the mRNA levels of NAMPT , as monitored by qRT ‐ PCR (left), and representative images from immunoblot assays against NAMPT are shown (right). The ratio of NAMPT to actin was quantified by densitometry, based on the immunoblot images from three independent experiments. (G) Cells were incubated with Met, BBR , and CC for 3 days. Representative images from immunoblots against NAMPT are shown. The ratio of NAMPT to β‐actin was quantified by densitometry based on the immunoblot images from three independent experiments. (H) H 2 O 2 ‐treated NIH 3T3 cells were incubated with NAMPT inhibitor ( FK 866, 5 nM ), QPRT inhibitor ( PHTH , 1 mM ), Met (10 mM ), and BBR (10 μM) alone or in combination as indicated for 3 days. The percentages of SA ‐β‐Gal‐positive cells are shown. (I) The mRNA levels of NMNAT 1 , NMNAT 2 , and NMNAT 3 genes in the cells infected with corresponding lentivirus‐sh RNA or nontargeting sh RNA (sh CON ) as determined by qRT ‐ PCR . (J) Cellular concentrations of NAD + in sh RNA ‐infected cells. (K) Cells infected with sh RNA s were treated with H 2 O 2 and incubated with BBR for 3 days. Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). * P

    Article Snippet: Quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed using SYBR Green Supermix kit (Bio‐Rad, CA, USA) on a Bio‐Rad IQ5 system.

    Techniques: Staining, Incubation, Quantitative RT-PCR, Western Blot, Infection

    AMPK restored the autophagic flux associated with the amelioration of lysosomal function, mTOR inactivation, but not the nuclear translocation of TFEB . A‐C: H 2 O 2 ‐treated NIH 3T3 cells were incubated in complete medium with different reagents for 3 days as indicated. (A) Representative images from immunoblot assays against Cathepsin B protein. (B) The acid phosphatase activity in cells. (C) Representative images from immunoblot assays against phosphorylated mTOR (pm TOR , Ser2448), phosphorylated 70S6K (P‐p70S6K, Thr389), pAMPK (Thr172), and β‐actin. (D) Immunofluorescent images of TFEB after treatment with indicated reagents. The bar represents 20 μm. (E) Immunoblot assays against TFEB protein of total cytoplasmic and nuclear subcellular fractions obtained from NIH 3T3 cells with the indicated treatment. (F) Relative fold‐changes in mRNA levels of two TFEB target genes ( GNS and LAMP ‐1 ) were monitored by qRT ‐ PCR assays.* P

    Journal: Aging Cell

    Article Title: AMPK activation protects cells from oxidative stress‐induced senescence via autophagic flux restoration and intracellular NAD+ elevation

    doi: 10.1111/acel.12446

    Figure Lengend Snippet: AMPK restored the autophagic flux associated with the amelioration of lysosomal function, mTOR inactivation, but not the nuclear translocation of TFEB . A‐C: H 2 O 2 ‐treated NIH 3T3 cells were incubated in complete medium with different reagents for 3 days as indicated. (A) Representative images from immunoblot assays against Cathepsin B protein. (B) The acid phosphatase activity in cells. (C) Representative images from immunoblot assays against phosphorylated mTOR (pm TOR , Ser2448), phosphorylated 70S6K (P‐p70S6K, Thr389), pAMPK (Thr172), and β‐actin. (D) Immunofluorescent images of TFEB after treatment with indicated reagents. The bar represents 20 μm. (E) Immunoblot assays against TFEB protein of total cytoplasmic and nuclear subcellular fractions obtained from NIH 3T3 cells with the indicated treatment. (F) Relative fold‐changes in mRNA levels of two TFEB target genes ( GNS and LAMP ‐1 ) were monitored by qRT ‐ PCR assays.* P

    Article Snippet: Quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed using SYBR Green Supermix kit (Bio‐Rad, CA, USA) on a Bio‐Rad IQ5 system.

    Techniques: Translocation Assay, Incubation, Activity Assay, Quantitative RT-PCR

    Activation of AMPK prevented H 2 O 2 ‐induced senescence. A to D: H 2 O 2 ‐treated NIH 3T3 cells were incubated in complete medium with metformin (Met, 5 to 10 mM ) or berberine ( BBR , 5 to 10 μM) for 3 days. (A) Representative images from immunoblot assays against pAMPK α (Thr172), AMPK α1, pACC (Ser79), and β‐actin. (B) Relative fold‐changes in mRNA levels of CPT ‐1 and FAS as determined by qRT ‐ PCR . (C) Representative images of SA ‐β‐Gal staining of cells (left), and percentages of SA ‐β‐Gal‐positive cells. D‐F: NIH 3T3 cells were treated with H 2 O 2 and incubated with Met (10 mM ), BBR (10 μM) and an AMPK inhibitor, Compound C ( CC , 10 μM), alone or in combination for 3 days. (D) Representative images from immunoblot assays. (E) Relative mRNA levels of CPT ‐1 and FAS as determined by qRT ‐ PCR . (F) Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). G‐I: NIH 3T3 cells without H 2 O 2 treatment were used. (G) The decrease in AMPK activity in DN ‐ AMPK ‐expressing NIH 3T3 cells was shown by the decrease in AMPK α phosphorylation. (H) Representative images of SA ‐β‐Gal staining of the nontransfected cells with or without CC (upper) and cells transfection with DN ‐ AMPK (lower). (I) The percentages of SA ‐β‐Gal‐positive cells were calculated based on the images represented in H. (J) H 2 O 2 ‐treated cells incubated with or without Met (10 mM ) or BBR (10 μM) for 3 days, representative images of SA ‐β‐Gal staining of cells transfected with empty vector (upper) or DN ‐ AMPK (lower). (K) The percentages of SA ‐β‐Gal‐positive cells were calculated based on the images presented in J. * P

    Journal: Aging Cell

    Article Title: AMPK activation protects cells from oxidative stress‐induced senescence via autophagic flux restoration and intracellular NAD+ elevation

    doi: 10.1111/acel.12446

    Figure Lengend Snippet: Activation of AMPK prevented H 2 O 2 ‐induced senescence. A to D: H 2 O 2 ‐treated NIH 3T3 cells were incubated in complete medium with metformin (Met, 5 to 10 mM ) or berberine ( BBR , 5 to 10 μM) for 3 days. (A) Representative images from immunoblot assays against pAMPK α (Thr172), AMPK α1, pACC (Ser79), and β‐actin. (B) Relative fold‐changes in mRNA levels of CPT ‐1 and FAS as determined by qRT ‐ PCR . (C) Representative images of SA ‐β‐Gal staining of cells (left), and percentages of SA ‐β‐Gal‐positive cells. D‐F: NIH 3T3 cells were treated with H 2 O 2 and incubated with Met (10 mM ), BBR (10 μM) and an AMPK inhibitor, Compound C ( CC , 10 μM), alone or in combination for 3 days. (D) Representative images from immunoblot assays. (E) Relative mRNA levels of CPT ‐1 and FAS as determined by qRT ‐ PCR . (F) Representative images of SA ‐β‐Gal staining of cells (left) and percentages of SA ‐β‐Gal‐positive cells (right). G‐I: NIH 3T3 cells without H 2 O 2 treatment were used. (G) The decrease in AMPK activity in DN ‐ AMPK ‐expressing NIH 3T3 cells was shown by the decrease in AMPK α phosphorylation. (H) Representative images of SA ‐β‐Gal staining of the nontransfected cells with or without CC (upper) and cells transfection with DN ‐ AMPK (lower). (I) The percentages of SA ‐β‐Gal‐positive cells were calculated based on the images represented in H. (J) H 2 O 2 ‐treated cells incubated with or without Met (10 mM ) or BBR (10 μM) for 3 days, representative images of SA ‐β‐Gal staining of cells transfected with empty vector (upper) or DN ‐ AMPK (lower). (K) The percentages of SA ‐β‐Gal‐positive cells were calculated based on the images presented in J. * P

    Article Snippet: Quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed using SYBR Green Supermix kit (Bio‐Rad, CA, USA) on a Bio‐Rad IQ5 system.

    Techniques: Activation Assay, Incubation, Cycling Probe Technology, Quantitative RT-PCR, Staining, Activity Assay, Expressing, Transfection, Plasmid Preparation

    H 2 O 2 induced senescence and AMPK pathway inhibition in NIH 3T3 Cells. Cells were treated with H 2 O 2 and incubated in complete medium without H 2 O 2 for 3‐5 days. CTL means untreated cells. (A) Representative images of SA ‐β‐Gal staining of the cells (left) and percentages of SA ‐β‐Gal‐positive cells in a total of 1000 cells (right). (B) Representative images of SAHF s in cells (left) and percentages of SAHF s‐positive cells in 1000 cells (right). (C) Representative images from immunoblot assays against p53 and β‐actin. (D) Relative fold‐changes in the mRNA levels of the genes encoding p21, IL 6 and IL 8 , as determined by qRT ‐ PCR . (E) Representative images from immunoblot assays against phosphorylated AMPK α ( pAMPK , Thr172), AMPK α1, phosphorylated ACC ( pACC , Ser79), and β‐actin. (F) The ratio of pAMPK to total AMPK was quantified by densitometry based on immunoblot images from three independent experiments. (G) Relative fold‐changes in the mRNA levels of two AMPK target genes ( CPT ‐1 and FAS ) were monitored by qRT ‐ PCR assays. * P

    Journal: Aging Cell

    Article Title: AMPK activation protects cells from oxidative stress‐induced senescence via autophagic flux restoration and intracellular NAD+ elevation

    doi: 10.1111/acel.12446

    Figure Lengend Snippet: H 2 O 2 induced senescence and AMPK pathway inhibition in NIH 3T3 Cells. Cells were treated with H 2 O 2 and incubated in complete medium without H 2 O 2 for 3‐5 days. CTL means untreated cells. (A) Representative images of SA ‐β‐Gal staining of the cells (left) and percentages of SA ‐β‐Gal‐positive cells in a total of 1000 cells (right). (B) Representative images of SAHF s in cells (left) and percentages of SAHF s‐positive cells in 1000 cells (right). (C) Representative images from immunoblot assays against p53 and β‐actin. (D) Relative fold‐changes in the mRNA levels of the genes encoding p21, IL 6 and IL 8 , as determined by qRT ‐ PCR . (E) Representative images from immunoblot assays against phosphorylated AMPK α ( pAMPK , Thr172), AMPK α1, phosphorylated ACC ( pACC , Ser79), and β‐actin. (F) The ratio of pAMPK to total AMPK was quantified by densitometry based on immunoblot images from three independent experiments. (G) Relative fold‐changes in the mRNA levels of two AMPK target genes ( CPT ‐1 and FAS ) were monitored by qRT ‐ PCR assays. * P

    Article Snippet: Quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed using SYBR Green Supermix kit (Bio‐Rad, CA, USA) on a Bio‐Rad IQ5 system.

    Techniques: Inhibition, Incubation, CTL Assay, Staining, Quantitative RT-PCR, Cycling Probe Technology

    Vitamin D3 (VitD3) upregulates endogenous osteopontin (OPN) -A and -C isomers via a VDR-dependent pathway after subarachnoid hemorrhage (SAH). (a) Effect of VDR siRNA on the VDR mRNA expression. Pre-SAH + VitD3, intranasal treatment 30 ng/kg/day VitD3 24 h before SAH; siRNA, small interfering ribonucleic acid. (b) Structure of OPN isomers and qRT-PCR universal primer design. (c) VitD3 upregulates endogenous OPN-A and -C isomers. * P

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Intranasal administration of vitamin D attenuates blood–brain barrier disruption through endogenous upregulation of osteopontin and activation of CD44/P-gp glycosylation signaling after subarachnoid hemorrhage in rats

    doi: 10.1177/0271678X16671147

    Figure Lengend Snippet: Vitamin D3 (VitD3) upregulates endogenous osteopontin (OPN) -A and -C isomers via a VDR-dependent pathway after subarachnoid hemorrhage (SAH). (a) Effect of VDR siRNA on the VDR mRNA expression. Pre-SAH + VitD3, intranasal treatment 30 ng/kg/day VitD3 24 h before SAH; siRNA, small interfering ribonucleic acid. (b) Structure of OPN isomers and qRT-PCR universal primer design. (c) VitD3 upregulates endogenous OPN-A and -C isomers. * P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) reactions were conducted using the SYBR Green detection reagent in Bio-Rad iQ5 system (Hercules, California).

    Techniques: Expressing, Quantitative RT-PCR

    Expression profile of selected miRNAs from qRT-PCR analysis

    Journal: BMC Plant Biology

    Article Title: Computational prediction of miRNAs and their targets in Phaseolus vulgaris using simple sequence repeat signatures

    doi: 10.1186/s12870-015-0516-3

    Figure Lengend Snippet: Expression profile of selected miRNAs from qRT-PCR analysis

    Article Snippet: Quantitative Real Time Polymerase Chain Reaction (qRT-PCR) qRT-PCR reactions were carried out for the five selected miRNAs in a Bio-Rad CFX96 Real-Time PCR system using Bio-Rad iQ SYBR green supermix.

    Techniques: Expressing, Quantitative RT-PCR

    qRT-PCR analysis of IL-17 and forkhead box P3 (Foxp3) in the spleen ( n = 3). Note that the Foxp3 level was increased, but the IL-17 level was similar in the spleen of the CAN group compared with the NC group. *** p

    Journal: Cell Transplantation

    Article Title: Differences in Tfh Cell Response between the Graft and Spleen with Chronic Allograft Nephropathy

    doi: 10.3727/096368916X692816

    Figure Lengend Snippet: qRT-PCR analysis of IL-17 and forkhead box P3 (Foxp3) in the spleen ( n = 3). Note that the Foxp3 level was increased, but the IL-17 level was similar in the spleen of the CAN group compared with the NC group. *** p

    Article Snippet: The mRNA levels were detected by quantitative realtime polymerase chain reaction (qRT-PCR) using the SYBR® Premix Ex TaqTM Kit (Takara) and the Prism 7500 RT-PCR System (Applied Biosystems, Foster City, CA, USA).

    Techniques: Quantitative RT-PCR

    Comparison of C-X-C chemokine receptor 5 (CXCR5) and B-cell-activating factor (BAFF) mRNA levels between the renal graft and spleen with CAN using qRT-PCR ( n = 3). Note that lower CXCR5 and BAFF levels were observed in the spleen than those in the graft of the CAN group. *** p

    Journal: Cell Transplantation

    Article Title: Differences in Tfh Cell Response between the Graft and Spleen with Chronic Allograft Nephropathy

    doi: 10.3727/096368916X692816

    Figure Lengend Snippet: Comparison of C-X-C chemokine receptor 5 (CXCR5) and B-cell-activating factor (BAFF) mRNA levels between the renal graft and spleen with CAN using qRT-PCR ( n = 3). Note that lower CXCR5 and BAFF levels were observed in the spleen than those in the graft of the CAN group. *** p

    Article Snippet: The mRNA levels were detected by quantitative realtime polymerase chain reaction (qRT-PCR) using the SYBR® Premix Ex TaqTM Kit (Takara) and the Prism 7500 RT-PCR System (Applied Biosystems, Foster City, CA, USA).

    Techniques: Quantitative RT-PCR

    FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by qRT-PCR. (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).

    Journal: Frontiers in Oncology

    Article Title: FA2H Exhibits Tumor Suppressive Roles on Breast Cancers via Cancer Stemness Control

    doi: 10.3389/fonc.2019.01089

    Figure Lengend Snippet: FA2H expression among breast cancer cell lines. (A) FA2H gene expression among different breast cancer cell lines by qRT-PCR. (B) FA2H protein expression among different breast cancer cell lines by Western-blotting. (C) FA2H protein expression quantified using the ImageJ software. (D) FA2H gene expression across 56 breast cancer cell lines ( 26 ).

    Article Snippet: Primers designed for candidate genes and used for quantitative reverse transcription polymerase chain reaction (qRT-PCR) validation (ordered from GENEWIZ) were listed in .

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software

    MYC3 and MYC4 regulate expression of JA-responsive genes. (A, B) qRT-PCR expression analysis of JAZ genes and wound-responsive genes in 10-d-old wild-type and MYC overexpression seedlings without JA treatment. Wild-type samples served as a calibrator for the calculation of relative expression levels (arbitrarily set to one). (C) Relative transcript levels of the PDF1.2 gene in 10-d-old wild-type and overexpression transgenic seedlings with or without 10 μM JA treatment (incubated for 6 h). Wild-type samples without JA treatment served as a calibrator, and relative expression was determined from replicate measurements in two independent biological replicates. Data are mean ±SE.

    Journal: Journal of Experimental Botany

    Article Title: Characterization of JAZ-interacting bHLH transcription factors that regulate jasmonate responses in Arabidopsis

    doi: 10.1093/jxb/erq408

    Figure Lengend Snippet: MYC3 and MYC4 regulate expression of JA-responsive genes. (A, B) qRT-PCR expression analysis of JAZ genes and wound-responsive genes in 10-d-old wild-type and MYC overexpression seedlings without JA treatment. Wild-type samples served as a calibrator for the calculation of relative expression levels (arbitrarily set to one). (C) Relative transcript levels of the PDF1.2 gene in 10-d-old wild-type and overexpression transgenic seedlings with or without 10 μM JA treatment (incubated for 6 h). Wild-type samples without JA treatment served as a calibrator, and relative expression was determined from replicate measurements in two independent biological replicates. Data are mean ±SE.

    Article Snippet: qRT-PCR analysis Polymerase chain reactions were performed with an Mx3005P Real-Time PCR System (Stratagene, CA), using SYBR® Green to monitor dsDNA synthesis.

    Techniques: Expressing, Quantitative RT-PCR, Over Expression, Transgenic Assay, Incubation

    PUM1 regulates ATXN1 levels via a highly conserved binding motif (A) Schematic representation of Human ATXN1 -3′UTR showing three putative PUM1 binding motifs (gray and red boxes) and their conservation in different species. The numbers indicate positions of PUM1 motifs in the human ATXN1 -3′UTR. (B) ATXN1 mRNA quantification by qRT-PCR in HEK293T cells upon overexpression (left panel) or knockdown (si PUM1 -81 and -82) (right panel) of PUM1 . The destination-cloning vector (Control), scrambled siRNAs (siScr.) and cel-miR-67 were used as negative controls. The housekeeping gene GAPDH was used to normalize the expression of genes in all the qRT-PCR experiments. (C) Luciferase assay in HEK293 cells overexpressing the reporter construct harboring the full length ATXN1 -3′UTR. In these conditions, we overexpressed (left panel) or decreased expression (right panel) of PUM1 . (D) Luciferase assay in HEK293T cells transfected with single WT and mutant (MUT) putative PUM1 binding site on the ATXN1 -3′UTR. The position of cloned regions for each PUM1 binding motif are indicated. The mutagenized nucleotides are highlighted in blue. In panels C and D the destination-vector (control), RNAi scramble (siScramble) and cel-miR-67 were used as negative controls; miR-101a was used as positive control. (RL) Renilla, (FL) Firefly luciferase. All the experiments were performed in triplicate (data represent mean ± S.E.M.). P values were calculated by Student’s t-test. Statistical significance is indicated as follows: * P

    Journal: Cell

    Article Title: Pumilio1 Haploinsufficiency Leads to SCA1-like Neurodegeneration by Increasing Wild-Type Ataxin1 Levels

    doi: 10.1016/j.cell.2015.02.012

    Figure Lengend Snippet: PUM1 regulates ATXN1 levels via a highly conserved binding motif (A) Schematic representation of Human ATXN1 -3′UTR showing three putative PUM1 binding motifs (gray and red boxes) and their conservation in different species. The numbers indicate positions of PUM1 motifs in the human ATXN1 -3′UTR. (B) ATXN1 mRNA quantification by qRT-PCR in HEK293T cells upon overexpression (left panel) or knockdown (si PUM1 -81 and -82) (right panel) of PUM1 . The destination-cloning vector (Control), scrambled siRNAs (siScr.) and cel-miR-67 were used as negative controls. The housekeeping gene GAPDH was used to normalize the expression of genes in all the qRT-PCR experiments. (C) Luciferase assay in HEK293 cells overexpressing the reporter construct harboring the full length ATXN1 -3′UTR. In these conditions, we overexpressed (left panel) or decreased expression (right panel) of PUM1 . (D) Luciferase assay in HEK293T cells transfected with single WT and mutant (MUT) putative PUM1 binding site on the ATXN1 -3′UTR. The position of cloned regions for each PUM1 binding motif are indicated. The mutagenized nucleotides are highlighted in blue. In panels C and D the destination-vector (control), RNAi scramble (siScramble) and cel-miR-67 were used as negative controls; miR-101a was used as positive control. (RL) Renilla, (FL) Firefly luciferase. All the experiments were performed in triplicate (data represent mean ± S.E.M.). P values were calculated by Student’s t-test. Statistical significance is indicated as follows: * P

    Article Snippet: Quantitative RT-polymerase chain reaction (qRT-PCR) experiments were performed using the CFX96 Touch Real-Time PCR Detection System (Bio-Rad Laboratories) with PerfeCta SYBR Green FastMix, ROX (Quanta Biosciences).

    Techniques: Binding Assay, Quantitative RT-PCR, Over Expression, Clone Assay, Plasmid Preparation, Expressing, Luciferase, Construct, Transfection, Mutagenesis, Positive Control

    ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by qRT-PCR (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.

    Journal: International Journal of Molecular Sciences

    Article Title: MicroRNA Expression Profile of Neural Progenitor-Like Cells Derived from Rat Bone Marrow Mesenchymal Stem Cells under the Influence of IGF-1, bFGF and EGF

    doi: 10.3390/ijms16059693

    Figure Lengend Snippet: ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by qRT-PCR (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.

    Article Snippet: MicroRNA-qPCR Assay MicroRNA expressions were validated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) according to the manufacturer’s protocol (all from Applied Biosystems, Life Technologies Co.; Waltham, MA, USA).

    Techniques: Negative Control, Expressing, Quantitative RT-PCR, Nucleic Acid Electrophoresis

    Characterization of apoptosis triggered by TCEA3 expression. a RH30 cells expressing TCEA3 or empty vector control were harvested for RNA. BAX and BCL2 gene expression were assayed by qRT-PCR. Scale bars 50 µm. b RH30 cells expressing TCEA3 or empty vector were assayed by western blot with antibodies against the indicated apoptotic marker proteins. GAPDH was used as a loading control. c RH30 cells expressing TCEA3 and empty vector were treated with caspase 8 inhibitor (Z-IETD-FMK, 40 μM), caspase 9 inhibitor (Z-LEHD-FMK, 40 μM) or a pan-caspase inhibitor (Z-VAD-FMK, 50 μM) for 18 h. Immunofluorescence assay was done with anti TCEA3 antibody (green) and anti-cleaved caspase 3 antibody (red). DAPI (blue) was used to visualize nuclei. Scale bar is 50 µm. d Western blot assay on the same cells as in C. to confirm the protein expression of TCEA3, caspase 3 and tubulin as a loading control. * marks cleaved caspase 3. Error bars are S.E.M. Student t test; *** p

    Journal: Cell Death & Disease

    Article Title: The transcription elongation factor TCEA3 induces apoptosis in rhabdomyosarcoma

    doi: 10.1038/s41419-020-2258-x

    Figure Lengend Snippet: Characterization of apoptosis triggered by TCEA3 expression. a RH30 cells expressing TCEA3 or empty vector control were harvested for RNA. BAX and BCL2 gene expression were assayed by qRT-PCR. Scale bars 50 µm. b RH30 cells expressing TCEA3 or empty vector were assayed by western blot with antibodies against the indicated apoptotic marker proteins. GAPDH was used as a loading control. c RH30 cells expressing TCEA3 and empty vector were treated with caspase 8 inhibitor (Z-IETD-FMK, 40 μM), caspase 9 inhibitor (Z-LEHD-FMK, 40 μM) or a pan-caspase inhibitor (Z-VAD-FMK, 50 μM) for 18 h. Immunofluorescence assay was done with anti TCEA3 antibody (green) and anti-cleaved caspase 3 antibody (red). DAPI (blue) was used to visualize nuclei. Scale bar is 50 µm. d Western blot assay on the same cells as in C. to confirm the protein expression of TCEA3, caspase 3 and tubulin as a loading control. * marks cleaved caspase 3. Error bars are S.E.M. Student t test; *** p

    Article Snippet: Two microgram of total RNA was reverse transcribed with MultiScribe TM MuLV reverse transcriptase (Life Technologies, Carlsbad, CA) and 40 ng of cDNA was used for quantitative real-time polymerase chain reaction (qRT-PCR) amplification (Life Technologies, Carlsbad, CA) with SYBR green PCR master mix (Life Technologies, Carlsbad, CA).

    Techniques: Expressing, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Marker, Immunofluorescence

    TCEA3 expression is downregulated in RMS. a TCEA3 mRNA expression was assayed in C2C12 cells, ARMS (RH28, RH30) and ERMS (RD, RD2) cell lines, respectively, by qRT-PCR. b Western blot with antibodies against TCEA3 to check expression in ERMS (RD, RD2), ARMS (RH28, RH30), and C2C12 cells. c TCEA3 immunostaining with antibodies against TCEA3 and DAPI was used to stain the nuclei. Scale bar is 100 µm. Error bars are S.E.M, Student's t test; *** p

    Journal: Cell Death & Disease

    Article Title: The transcription elongation factor TCEA3 induces apoptosis in rhabdomyosarcoma

    doi: 10.1038/s41419-020-2258-x

    Figure Lengend Snippet: TCEA3 expression is downregulated in RMS. a TCEA3 mRNA expression was assayed in C2C12 cells, ARMS (RH28, RH30) and ERMS (RD, RD2) cell lines, respectively, by qRT-PCR. b Western blot with antibodies against TCEA3 to check expression in ERMS (RD, RD2), ARMS (RH28, RH30), and C2C12 cells. c TCEA3 immunostaining with antibodies against TCEA3 and DAPI was used to stain the nuclei. Scale bar is 100 µm. Error bars are S.E.M, Student's t test; *** p

    Article Snippet: Two microgram of total RNA was reverse transcribed with MultiScribe TM MuLV reverse transcriptase (Life Technologies, Carlsbad, CA) and 40 ng of cDNA was used for quantitative real-time polymerase chain reaction (qRT-PCR) amplification (Life Technologies, Carlsbad, CA) with SYBR green PCR master mix (Life Technologies, Carlsbad, CA).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunostaining, Staining

    TCEA3 is repressed by TBX2 and expression correlates with DNA methylation. a RH30 cells depleted for TBX2 or scr control were assayed for mRNA expression of TCEA3 by qRT-PCR. b Chromatin immunoprecipitation (ChIP) assays with antibodies against TBX2 and IgG were assayed with primers against the TCEA3 promoter. c Scatterplot for TCEA3 mRNA expression (RNAseq) and TCEA3 locus methylation level (RRBS) was generated for 834 cancer cell lines using the Cancer Cell Line Encyclopedia (CCLE) database, and the plot was edited using Plotly tool. Each dot represents a cell line. mRNA expression value below 0 have no or negligible expression, and the DNA methylation levels values ranging from 0 to 1 denoting unmethylated to fully methylated locus, respectively. d Inhibition of DNA methylation derepresses TCEA3 expression in RH30 cells. mRNA expression of TCEA3 and TCEA1 were assayed in RH30 cells treated with either DMSO vehicle control or 5-aza-2’deoxycytidine DNA methyltransferase inhibitor at 30 μM and 60 μM concentrations for 48 h by qRT-PCR. Error bars are S.E.M. Student's t test; ns represents ‘not significant’, *** p

    Journal: Cell Death & Disease

    Article Title: The transcription elongation factor TCEA3 induces apoptosis in rhabdomyosarcoma

    doi: 10.1038/s41419-020-2258-x

    Figure Lengend Snippet: TCEA3 is repressed by TBX2 and expression correlates with DNA methylation. a RH30 cells depleted for TBX2 or scr control were assayed for mRNA expression of TCEA3 by qRT-PCR. b Chromatin immunoprecipitation (ChIP) assays with antibodies against TBX2 and IgG were assayed with primers against the TCEA3 promoter. c Scatterplot for TCEA3 mRNA expression (RNAseq) and TCEA3 locus methylation level (RRBS) was generated for 834 cancer cell lines using the Cancer Cell Line Encyclopedia (CCLE) database, and the plot was edited using Plotly tool. Each dot represents a cell line. mRNA expression value below 0 have no or negligible expression, and the DNA methylation levels values ranging from 0 to 1 denoting unmethylated to fully methylated locus, respectively. d Inhibition of DNA methylation derepresses TCEA3 expression in RH30 cells. mRNA expression of TCEA3 and TCEA1 were assayed in RH30 cells treated with either DMSO vehicle control or 5-aza-2’deoxycytidine DNA methyltransferase inhibitor at 30 μM and 60 μM concentrations for 48 h by qRT-PCR. Error bars are S.E.M. Student's t test; ns represents ‘not significant’, *** p

    Article Snippet: Two microgram of total RNA was reverse transcribed with MultiScribe TM MuLV reverse transcriptase (Life Technologies, Carlsbad, CA) and 40 ng of cDNA was used for quantitative real-time polymerase chain reaction (qRT-PCR) amplification (Life Technologies, Carlsbad, CA) with SYBR green PCR master mix (Life Technologies, Carlsbad, CA).

    Techniques: Expressing, DNA Methylation Assay, Quantitative RT-PCR, Chromatin Immunoprecipitation, Methylation, Generated, Inhibition

    Exogenous TCEA3 overexpression in RMS cell lines. a RH30, RH28, RD, and RD2 cell lines were transfected with a TCEA3-expressiion plasmid (pTCEA3) or the empty vector (EV). TCEA3 ectopic expression was confirmed by assaying TCEA3 mRNA expression by qRT-PCR. b Western blot with antibodies against TCEA3 was performed on the cells described in ( a ). to confirm the protein expression of TCEA3. GAPDH was used as the loading control. c TCEA3 immunofluorescence with antibodies against TCEA3 and DAPI used to stain nuclei on cells described in ( a ). Scale bar is 100 µm. Error bars are S.E.M. Student's t test; *** p

    Journal: Cell Death & Disease

    Article Title: The transcription elongation factor TCEA3 induces apoptosis in rhabdomyosarcoma

    doi: 10.1038/s41419-020-2258-x

    Figure Lengend Snippet: Exogenous TCEA3 overexpression in RMS cell lines. a RH30, RH28, RD, and RD2 cell lines were transfected with a TCEA3-expressiion plasmid (pTCEA3) or the empty vector (EV). TCEA3 ectopic expression was confirmed by assaying TCEA3 mRNA expression by qRT-PCR. b Western blot with antibodies against TCEA3 was performed on the cells described in ( a ). to confirm the protein expression of TCEA3. GAPDH was used as the loading control. c TCEA3 immunofluorescence with antibodies against TCEA3 and DAPI used to stain nuclei on cells described in ( a ). Scale bar is 100 µm. Error bars are S.E.M. Student's t test; *** p

    Article Snippet: Two microgram of total RNA was reverse transcribed with MultiScribe TM MuLV reverse transcriptase (Life Technologies, Carlsbad, CA) and 40 ng of cDNA was used for quantitative real-time polymerase chain reaction (qRT-PCR) amplification (Life Technologies, Carlsbad, CA) with SYBR green PCR master mix (Life Technologies, Carlsbad, CA).

    Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

    Pluripotency of in vivo rPSCs. ( A ) The established in vivo rPSCs expressed Oct4 -GFP and were morphologically indistinguishable from mESCs. Scale bar = 100 μm. ( B ) Immunocytochemistry analysis (scale bar = 50 μm) and ( C ) qRT-PCR showed that in vivo rPSCs expressed pluripotency markers Oct4 (endo), Nanog (endo), Sox2 , and Rex1 , similar to in vitro iPSCs and ESCs. ( D ) DNA methylation status of Oct4 and Nanog promoter region in rOG2-T-rPSCs #1, #2. ( E,F ) The rOG2-T-rPSCs could differentiate into all 3 germ layers in vitro and in vivo Scale bar = 100 μm. ( G ) Aggregation potential of in viv o rPSCs into the inner cell mass of normal embryos. The in vivo rPSCs did not contribute to the trophectoderm lineage. Scale bar = 50 μm.

    Journal: Scientific Reports

    Article Title: In vivo reprogrammed pluripotent stem cells from teratomas share analogous properties with their in vitro counterparts

    doi: 10.1038/srep13559

    Figure Lengend Snippet: Pluripotency of in vivo rPSCs. ( A ) The established in vivo rPSCs expressed Oct4 -GFP and were morphologically indistinguishable from mESCs. Scale bar = 100 μm. ( B ) Immunocytochemistry analysis (scale bar = 50 μm) and ( C ) qRT-PCR showed that in vivo rPSCs expressed pluripotency markers Oct4 (endo), Nanog (endo), Sox2 , and Rex1 , similar to in vitro iPSCs and ESCs. ( D ) DNA methylation status of Oct4 and Nanog promoter region in rOG2-T-rPSCs #1, #2. ( E,F ) The rOG2-T-rPSCs could differentiate into all 3 germ layers in vitro and in vivo Scale bar = 100 μm. ( G ) Aggregation potential of in viv o rPSCs into the inner cell mass of normal embryos. The in vivo rPSCs did not contribute to the trophectoderm lineage. Scale bar = 50 μm.

    Article Snippet: Quantitative polymerase chain reaction (qRT-PCR) reactions were set up in duplicate with the Power SYBR Green Master Mix (Takara) and analyzed with the Roche LightCycler 5480 (Roche).

    Techniques: In Vivo, Immunocytochemistry, Quantitative RT-PCR, In Vitro, DNA Methylation Assay

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

    Journal: Arthritis Research & Therapy

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

    doi: 10.1186/ar2109

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

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

    Techniques: Quantitative RT-PCR, Expressing, Amplification

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

    Journal: Arthritis Research & Therapy

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

    doi: 10.1186/ar2109

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

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

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

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

    Journal: Arthritis Research & Therapy

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

    doi: 10.1186/ar2109

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

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

    Techniques: Quantitative RT-PCR, Expressing, Amplification

    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, qRT‐PCR result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, qRT‐PCR result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Inhibition, Migration, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, EdU Assay, Transwell Assay

    MYC activated LINC01116 at transcriptional level in NPC. A‐D, The expression of MYC or LINC01116 in indicated CNE2 and 5‐8F cells was tested by qRT‐PCR. E, ChIP assay and AGE analysis verified the binding of MYC protein to LINC01116 promoter. F,G, Luciferase reporter assay confirmed that LINC01116 was transcriptionally activated by MYC in both CNE2 and 5‐8F cells. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: MYC activated LINC01116 at transcriptional level in NPC. A‐D, The expression of MYC or LINC01116 in indicated CNE2 and 5‐8F cells was tested by qRT‐PCR. E, ChIP assay and AGE analysis verified the binding of MYC protein to LINC01116 promoter. F,G, Luciferase reporter assay confirmed that LINC01116 was transcriptionally activated by MYC in both CNE2 and 5‐8F cells. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Binding Assay, Luciferase, Reporter Assay

    LINC01116 mainly located in the cytoplasm of NPC cells and interacted with the 5ʹUTR of MYC mRNA. A, LINC01116 was predicted by lncLocator as a cytoplasmic lncRNA. B, Subcellular fractionation plus qRT‐PCR validated that LINC01116 was mainly distributed in the cytoplasm of NPC cells. C, The online IntaRNA tool suggested that LINC01116 could bind to the 5'UTR of MYC mRNA. D, The interaction between LINC01116 and MYC mRNA in CNE2 and 5‐8F cells was confirmed by RNA pull down assay. ** P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: LINC01116 mainly located in the cytoplasm of NPC cells and interacted with the 5ʹUTR of MYC mRNA. A, LINC01116 was predicted by lncLocator as a cytoplasmic lncRNA. B, Subcellular fractionation plus qRT‐PCR validated that LINC01116 was mainly distributed in the cytoplasm of NPC cells. C, The online IntaRNA tool suggested that LINC01116 could bind to the 5'UTR of MYC mRNA. D, The interaction between LINC01116 and MYC mRNA in CNE2 and 5‐8F cells was confirmed by RNA pull down assay. ** P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Fractionation, Quantitative RT-PCR, Pull Down Assay

    LINC01116 enhanced MYC protein level so as to promote the transcription of MYC targets. A,B, The impact of LINC01116 on the mRNA and protein levels of MYC was estimated by qRT‐PCR (A) and western blot (B). C, Changes on the transcription of three MYC targets (CDK4, CCND2, and BMI1) in the context of MYC overexpression or together with LINC01116 suppression were evaluated by luciferase reporter assays. D‐E, qRT‐PCR results of the expression of CDK4, CCND2, and BMI1 in LINC01116‐silenced NPC cells. F, The influence of LINC01116 per se on the transcription of CDK4, CCND2, and BMI1 in NPC cells was determined by luciferase reporter assay. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: LINC01116 enhanced MYC protein level so as to promote the transcription of MYC targets. A,B, The impact of LINC01116 on the mRNA and protein levels of MYC was estimated by qRT‐PCR (A) and western blot (B). C, Changes on the transcription of three MYC targets (CDK4, CCND2, and BMI1) in the context of MYC overexpression or together with LINC01116 suppression were evaluated by luciferase reporter assays. D‐E, qRT‐PCR results of the expression of CDK4, CCND2, and BMI1 in LINC01116‐silenced NPC cells. F, The influence of LINC01116 per se on the transcription of CDK4, CCND2, and BMI1 in NPC cells was determined by luciferase reporter assay. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Quantitative RT-PCR, Western Blot, Over Expression, Luciferase, Expressing, Reporter Assay

    Overexpression of MYC rescued LINC01116 depletion‐restrained proliferation and migration in NPC cells. A, The expression of MYC at both mRNA and protein levels in indicated cells was detected via qRT‐PCR and western blot. B‐D, The viability, proliferation, and migration in 5‐8F cells under different conditions were respectively evaluated through CCK‐8 (B), EdU (C), and transwell assays (D). * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: Overexpression of MYC rescued LINC01116 depletion‐restrained proliferation and migration in NPC cells. A, The expression of MYC at both mRNA and protein levels in indicated cells was detected via qRT‐PCR and western blot. B‐D, The viability, proliferation, and migration in 5‐8F cells under different conditions were respectively evaluated through CCK‐8 (B), EdU (C), and transwell assays (D). * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay

    Genome-wide analysis for cryptic SVs identifies disruption of further neurodevelopmental disease candidate genes and demonstrates reduced expression of ZNF804A in patient cells. (A,B) Graphical representation of the three genes disrupted by an ∼10 kB deletion on chr 19 [del(19q13.4)] (A) and the cryptic paracentric inversion inv(2)(p32.1q32.1) in the patient. Sites of breakpoints are denoted by arrows. (C) Graphical representation of anomalous-read (red dots) fusion positions for the cryptic 2.49 Mb paracentric inversion on chromosome 2. Mate-pair library sequencing-predicted breakpoints 5′ and 3′ of the inversion were amplified with breakpoint-specific primers and validated at base-pair level by PCR and capillary sequencing. (D) mRNA-levels of ZNF804A and the housekeeping gene RPL19 were quantified by qRT-PCR from total RNA isolated from fibroblasts of the patient or a healthy male control and normalized to expression of beta-actin.

    Journal: PLoS ONE

    Article Title: Sequencing of a Patient with Balanced Chromosome Abnormalities and Neurodevelopmental Disease Identifies Disruption of Multiple High Risk Loci by Structural Variation

    doi: 10.1371/journal.pone.0090894

    Figure Lengend Snippet: Genome-wide analysis for cryptic SVs identifies disruption of further neurodevelopmental disease candidate genes and demonstrates reduced expression of ZNF804A in patient cells. (A,B) Graphical representation of the three genes disrupted by an ∼10 kB deletion on chr 19 [del(19q13.4)] (A) and the cryptic paracentric inversion inv(2)(p32.1q32.1) in the patient. Sites of breakpoints are denoted by arrows. (C) Graphical representation of anomalous-read (red dots) fusion positions for the cryptic 2.49 Mb paracentric inversion on chromosome 2. Mate-pair library sequencing-predicted breakpoints 5′ and 3′ of the inversion were amplified with breakpoint-specific primers and validated at base-pair level by PCR and capillary sequencing. (D) mRNA-levels of ZNF804A and the housekeeping gene RPL19 were quantified by qRT-PCR from total RNA isolated from fibroblasts of the patient or a healthy male control and normalized to expression of beta-actin.

    Article Snippet: Presence of abnormal gene products was validated by amplifying proposed fusion mRNAs isolated from patient and control lymphoblasts by quantitative-reverse polymerase chain reaction (qRT-PCR) using SYBR Green Supermix (Bio-Rad, Hercules, CA) according to established protocols.

    Techniques: Genome Wide, Expressing, Sequencing, Amplification, Polymerase Chain Reaction, Quantitative RT-PCR, Isolation

    Analysis of HIC1 expression in colorectal neoplasia. (A) HIC1 expression changes during tumor progression. qRT-PCR analysis of the HIC1 mRNA levels in healthy tissue, hyperplastic adenomas (Hyp; n = 9), adenomas displaying low-grade (LGD; n = 24) or high-grade (HGD; n = 25) dysplasia, and CRC ( n = 12). The boxed areas correspond to the second and third quartiles; the median of ΔCp values for each category is indicated as the red line. The relation between the HIC1 expression profile and neoplasia progression is significant, as evidenced by the Spearman’s ( ρ = − 0.67) and Kendall’s ( τ = − 0.51) coefficient values. (B) HIC1 expression in CRC subgroups clustered according to the DNA methylation profiles. HIC1-high and HIC1-low samples are indicated by brown or blue dots, respectively. Black dots indicate the other specimens. The red lines correspond to the median values; log2-expression intensity, binary logarithm of expression intensity (additional details are given in Supplementary Methods).

    Journal: Translational Oncology

    Article Title: HIC1 Expression Distinguishes Intestinal Carcinomas Sensitive to Chemotherapy

    doi: 10.1016/j.tranon.2016.01.005

    Figure Lengend Snippet: Analysis of HIC1 expression in colorectal neoplasia. (A) HIC1 expression changes during tumor progression. qRT-PCR analysis of the HIC1 mRNA levels in healthy tissue, hyperplastic adenomas (Hyp; n = 9), adenomas displaying low-grade (LGD; n = 24) or high-grade (HGD; n = 25) dysplasia, and CRC ( n = 12). The boxed areas correspond to the second and third quartiles; the median of ΔCp values for each category is indicated as the red line. The relation between the HIC1 expression profile and neoplasia progression is significant, as evidenced by the Spearman’s ( ρ = − 0.67) and Kendall’s ( τ = − 0.51) coefficient values. (B) HIC1 expression in CRC subgroups clustered according to the DNA methylation profiles. HIC1-high and HIC1-low samples are indicated by brown or blue dots, respectively. Black dots indicate the other specimens. The red lines correspond to the median values; log2-expression intensity, binary logarithm of expression intensity (additional details are given in Supplementary Methods).

    Article Snippet: Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) Reactions were run in triplicates using LightCycler 480 Probes Master and Universal ProbeLibrary hydrolysis probes and LightCycler 480 Instrument (Roche Life Sciences).

    Techniques: Expressing, Quantitative RT-PCR, DNA Methylation Assay