for qrt pcr Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher qrt polymerase chain reaction pcr
    Qrt Polymerase Chain Reaction Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, 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 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    qrt polymerase chain reaction pcr - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    90
    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: 90/100, 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 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    transcription polymerase chain reaction qrt pcr system - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    91
    Thermo Fisher reverse transcription polymerase chain reaction qrt pcr
    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
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 583 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 91 stars, based on 583 article reviews
    Price from $9.99 to $1999.99
    reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    90
    Thermo Fisher reverse transcriptase polymerase chain reaction qrt pcr
    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
    Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 223 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 90 stars, based on 223 article reviews
    Price from $9.99 to $1999.99
    reverse transcriptase polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    91
    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: 91/100, based on 233 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 91 stars, based on 233 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 192 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 92 stars, based on 192 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    94
    Thermo Fisher polymerase chain reaction qrt pcr rneasy mini kit
    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
    Polymerase Chain Reaction Qrt Pcr Rneasy Mini Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 4 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 94 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr rneasy mini kit - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 1147 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 92 stars, based on 1147 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    88
    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: 88/100, based on 15 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 88 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    93
    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: 93/100, based on 383 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 93 stars, based on 383 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    89
    Roche 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
    Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction qrt pcr/product/Roche
    Average 89 stars, based on 36 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    99
    Thermo Fisher transcript polymerase chain reaction qrt pcr kit
    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
    Transcript Polymerase Chain Reaction Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, 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 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    transcript polymerase chain reaction qrt pcr kit - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    94
    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: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time polymerase chain reaction qrt pcr amplification/product/Thermo Fisher
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    real time polymerase chain reaction qrt pcr amplification - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    89
    Roche transcription polymerase chain reaction qrt pcr
    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
    Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 30 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 30 article reviews
    Price from $9.99 to $1999.99
    transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    91
    Agilent technologies quantitative real time polymerase chain reaction qrt pcr
    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
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, 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 91 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    94
    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: 94/100, based on 1503 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 94 stars, based on 1503 article reviews
    Price from $9.99 to $1999.99
    quantitative reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    94
    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: 94/100, based on 678 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 94 stars, based on 678 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    93
    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: 93/100, 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 93 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    reverse transcription polymerase chain reaction qrt pcr validation - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    94
    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: 94/100, based on 2668 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 94 stars, based on 2668 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    85
    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: 85/100, based on 17 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 85 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    qrt pcr analysis polymerase chain reactions - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    89
    Bio-Rad quantitative rt polymerase chain reaction qrt pcr 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.
    Quantitative Rt Polymerase Chain Reaction Qrt Pcr Reactions, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 55 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 89 stars, based on 55 article reviews
    Price from $9.99 to $1999.99
    quantitative rt polymerase chain reaction qrt pcr reactions - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    89
    Roche 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 Roche, used in various techniques. Bioz Stars score: 89/100, based on 62 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 62 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction qrt pcr analysis - by Bioz Stars, 2020-07
    89/100 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

    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

    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

    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

    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

    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

    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

    ( 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

    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

    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

    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

    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