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  • 99
    Thermo Fisher reverse transcription polymerase chain reaction rt pcr analysis total rna
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 280 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa reverse transcription polymerase chain reaction rt pcr analysis
    <t>Pluripotency</t> of patient-derived iPSCs. (A) Embryoid bodies expressed differentiation markers specific to all three germ layers. To examine the differentiation potential of iPSCs, we evaluated their ability to form embryoid bodies by using floating culture assays. <t>RT-PCR</t> was used to examine the expression of differentiation markers in the embryoid bodies. AFP was used as an endodermal marker, MSX1 as a mesodermal marker, and MAP as an ectodermal marker, with β-actin used as an internal control. (B) Teratoma formation in G-OFiPSCs. Patient-derived iPSCs (G-OF1iPS clones 1 and 6, and G-OF2iPS clones 5 and 8) were injected at a concentration of 1 × 10 6 cells in 20 μl of PBS into 8–10-week-old male CB17 SCID mice. Tumors were collected 9–12 weeks after the injections. Histological examination demonstrated that tissues originating from all three embryonic germ layers were present, including cartilage (mesoderm), gut-like epithelium (endoderm), and neural tube-like structures (ectoderm).
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Research Center inc reverse transcription polymerase chain reaction rt pcr analysis trizol
    <t>Pluripotency</t> of patient-derived iPSCs. (A) Embryoid bodies expressed differentiation markers specific to all three germ layers. To examine the differentiation potential of iPSCs, we evaluated their ability to form embryoid bodies by using floating culture assays. <t>RT-PCR</t> was used to examine the expression of differentiation markers in the embryoid bodies. AFP was used as an endodermal marker, MSX1 as a mesodermal marker, and MAP as an ectodermal marker, with β-actin used as an internal control. (B) Teratoma formation in G-OFiPSCs. Patient-derived iPSCs (G-OF1iPS clones 1 and 6, and G-OF2iPS clones 5 and 8) were injected at a concentration of 1 × 10 6 cells in 20 μl of PBS into 8–10-week-old male CB17 SCID mice. Tumors were collected 9–12 weeks after the injections. Histological examination demonstrated that tissues originating from all three embryonic germ layers were present, including cartilage (mesoderm), gut-like epithelium (endoderm), and neural tube-like structures (ectoderm).
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis Trizol, supplied by Molecular Research Center inc, used in various techniques. Bioz Stars score: 85/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad reverse transcription polymerase chain reaction rt pcr analysis
    Depletion of ChREBPβ attenuates glucose-stimulated β-cell proliferation in isolated rat islet cells. A and B : Isolated rat islet cells were treated with lipid-conjugated (Accell) siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h. Total <t>RNA</t> was collected and subjected to <t>RT-PCR</t> using primers specific for ChREBPβ or ChREBPα mRNAs. C : Protein extracts were subjected to immunoblotting using an antibody against ChREBPα and β-actin. D : To determine the effects of the siRNAs on ChREBP translocation, cells were treated with control or Accell siRNA against ChREBPβ cultured in 5.5 or 15 mmol/L glucose for 32 h and were fixed and stained for insulin, ChREBP, and DNA (DAPI). E : Isolated rat islet cells were treated with Accell siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h and fixed and stained with Ki67 and insulin. F : Quantification of the results in E , wherein at least 1,000 insulin-positive cells were counted. Results are from four different rat islet isolations. siChREBPβ-01 and -02, siRNAs directed against ChREBPβ; siCon, scramble control siRNA. Error bars are the SEM. * P
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche reverse transcription polymerase chain reaction rt pcr analysis
    Map-based cloning of the <t>z3</t> locus. a Physical mapping of the z3 locus. The z3 : Table S1. b Fine mapping of the z3 locus. The locus was further mapped within a 78-kb region between two markers, dCAPS7 and dCAPS2. Numbers below the line indicate F 2 recombinants at the marker regions. c Candidate genes (black boxes) in the 78-kb region. d T to C substitution of Z3 in the z3 mutant. Three exons and two introns are indicated as rectangles and lines, respectively. The position of the point mutation in the z3 mutant is at the third exon and represented by a black arrowhead. e Derived cleaved amplified polymorphic sequence (dCAPS) analysis of the point mutation in the z3 mutant. Aat ll was able to digest the genomic <t>PCR</t> products amplified from the WT, but not from the z3 mutant because of its T to C substitution in the Aat ll restriction enzyme site. M23, a mapping parent ‘Milyang23’; F1, F 1 hybrid ( z3 /Milyang23). f Z3 protein sequence. The predicted N-terminal signal peptide is marked with an open rectangle and the nine transmembrane domains (TMDs) are underlined. The yellow highlight represents regions predicted to be a CitMHS family. The position of the point mutation in the z3 mutant is highlighted in red
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Research Center inc reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Molecular Research Center inc, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman real time reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Taqman Real Time Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, 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
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    TaKaRa quantitative reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Quantitative Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies quantitative reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Quantitative Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene quantitative reverse transcription polymerase chain reaction rt pcr analysis
    <t>MUC5AC</t> gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by <t>RT-PCR</t> (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p
    Quantitative Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Stratagene, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rt pcr analysis
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Rt Pcr Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8597 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Meridian Life Science rt pcr analysis
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Rt Pcr Analysis, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 92/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcription polymerase chain reaction rt pcr analysis trizol reagent
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pasteur Institute rt pcr analysis
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Rt Pcr Analysis, supplied by Pasteur Institute, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher semiquantitative reverse transcription polymerase chain reaction rt pcr analysis total cellular rna
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Semiquantitative Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis Total Cellular Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co reverse transcription polymerase chain reaction rt pcr analysis total rna
    Effects of DIM and Herceptin on <t>miR-200</t> family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA <t>RT-PCR.</t> ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis Total Rna, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 91/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa real time reverse transcription polymerase chain reaction rt pcr analysis
    NRAS is a target of <t>miR-29a.</t> A, Sequence of the miR-29a binding site within the human NRAS 3′-UTR and a schematic diagram of the reporter construct showing the entire NRAS 3′-UTR sequence and the mutant NRAS 3′-UTR sequence. The mutant nucleotides of the NRAS 3′-UTR are labeled in red. B, Luciferase assay on H1299 cells which were cotransfected with miR-NC or miR-29a and a luciferase reporter containing the full length of NRAS 3′-UTR (WT) or a mutant type (MT) harboring 4 mutant nucleotides of the miR-29a binding site. Luciferase activities were measured 24 hours posttransfection. MicroRNA-29a markedly suppressed luciferase activity in NRAS 3′-UTR wild type (WT) reporter constructs. C, The immunoblotting showed that expression levels of NRAS were decreased in cells with miR-29a overexpression. D, The expression of NRAS in adjacent normal tissues and human lung cancer specimens was determined by quantitative <t>RT-PCR</t> analysis, and fold changes were obtained from the ratio of NRAS to GAPDH levels. E, Spearman rank correlation analysis showed that the expression levels of NRAS and miR-29a in lung cancer tissues were inversely correlated. Data represent mean ± SD of three replicates. **Significant difference at P
    Real Time Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad real time reverse transcription polymerase chain reaction rt pcr analysis
    NRAS is a target of <t>miR-29a.</t> A, Sequence of the miR-29a binding site within the human NRAS 3′-UTR and a schematic diagram of the reporter construct showing the entire NRAS 3′-UTR sequence and the mutant NRAS 3′-UTR sequence. The mutant nucleotides of the NRAS 3′-UTR are labeled in red. B, Luciferase assay on H1299 cells which were cotransfected with miR-NC or miR-29a and a luciferase reporter containing the full length of NRAS 3′-UTR (WT) or a mutant type (MT) harboring 4 mutant nucleotides of the miR-29a binding site. Luciferase activities were measured 24 hours posttransfection. MicroRNA-29a markedly suppressed luciferase activity in NRAS 3′-UTR wild type (WT) reporter constructs. C, The immunoblotting showed that expression levels of NRAS were decreased in cells with miR-29a overexpression. D, The expression of NRAS in adjacent normal tissues and human lung cancer specimens was determined by quantitative <t>RT-PCR</t> analysis, and fold changes were obtained from the ratio of NRAS to GAPDH levels. E, Spearman rank correlation analysis showed that the expression levels of NRAS and miR-29a in lung cancer tissues were inversely correlated. Data represent mean ± SD of three replicates. **Significant difference at P
    Real Time Reverse Transcription Polymerase Chain Reaction Rt Pcr Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Pluripotency of patient-derived iPSCs. (A) Embryoid bodies expressed differentiation markers specific to all three germ layers. To examine the differentiation potential of iPSCs, we evaluated their ability to form embryoid bodies by using floating culture assays. RT-PCR was used to examine the expression of differentiation markers in the embryoid bodies. AFP was used as an endodermal marker, MSX1 as a mesodermal marker, and MAP as an ectodermal marker, with β-actin used as an internal control. (B) Teratoma formation in G-OFiPSCs. Patient-derived iPSCs (G-OF1iPS clones 1 and 6, and G-OF2iPS clones 5 and 8) were injected at a concentration of 1 × 10 6 cells in 20 μl of PBS into 8–10-week-old male CB17 SCID mice. Tumors were collected 9–12 weeks after the injections. Histological examination demonstrated that tissues originating from all three embryonic germ layers were present, including cartilage (mesoderm), gut-like epithelium (endoderm), and neural tube-like structures (ectoderm).

    Journal: PLoS ONE

    Article Title: Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction

    doi: 10.1371/journal.pone.0186879

    Figure Lengend Snippet: Pluripotency of patient-derived iPSCs. (A) Embryoid bodies expressed differentiation markers specific to all three germ layers. To examine the differentiation potential of iPSCs, we evaluated their ability to form embryoid bodies by using floating culture assays. RT-PCR was used to examine the expression of differentiation markers in the embryoid bodies. AFP was used as an endodermal marker, MSX1 as a mesodermal marker, and MAP as an ectodermal marker, with β-actin used as an internal control. (B) Teratoma formation in G-OFiPSCs. Patient-derived iPSCs (G-OF1iPS clones 1 and 6, and G-OF2iPS clones 5 and 8) were injected at a concentration of 1 × 10 6 cells in 20 μl of PBS into 8–10-week-old male CB17 SCID mice. Tumors were collected 9–12 weeks after the injections. Histological examination demonstrated that tissues originating from all three embryonic germ layers were present, including cartilage (mesoderm), gut-like epithelium (endoderm), and neural tube-like structures (ectoderm).

    Article Snippet: The stemness and pluripotency of the iPSCs were analyzed using reverse transcription polymerase chain reaction (RT-PCR) analysis with ExTaq DNA polymerase (Takara Biotechnology, Shiga, Japan).

    Techniques: Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Marker, Clone Assay, Injection, Concentration Assay, Mouse Assay

    Hh-dependent induction of osteogenic marker ALP in patient-derived iPSCs. (A) Control iPSCs (KD iPSCs, 201B7iPS cells, and Nips B2 iPSCs) and patient-derived iPSCs (G-OF1iPS1, and G-OF2iPS5) were cultured in OBM, OBM with FIT (bFGF, IGF-1, and TGF-β1), or OBM with SAG. FIT- and SAG-induced ALP activity significantly increased in patient-derived iPSCs compared with control iPSCs. (B) A sample of each supernatant was transferred to tubes for measurement of ALP activity as the hydrolysis of ρ-nitrophenyl phosphate using a LabAssay ALP kit and protein concentration using a Protein assay rapid kit. G-OF1iPS1 was showed a high ALP activity compared with 201B7,especially cultured in OBM with SAG. (C) KDiPSCs and G-OF1iPS1 cells were cultured in OBM (control), or OBM supplemented with 100 ng/ml BMP2/7 for 24 h. qRT-PCR analysis of RUNX2 was conducted using Premix Ex Taq reagent according to the manufacturer’s instructions. RUNX2 levels were normalized to those of GAPDH. A significant difference was observed in G-OF1iPS1. Especially, BMP2/7-induced RUNX2 expression promoted an approximate 2-fold increase in the number of G-OFiPSCs compared with KDiPSCs. Values represent the mean ± SD. Bonferroni correction for multiple comparisons was applied, * p

    Journal: PLoS ONE

    Article Title: Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction

    doi: 10.1371/journal.pone.0186879

    Figure Lengend Snippet: Hh-dependent induction of osteogenic marker ALP in patient-derived iPSCs. (A) Control iPSCs (KD iPSCs, 201B7iPS cells, and Nips B2 iPSCs) and patient-derived iPSCs (G-OF1iPS1, and G-OF2iPS5) were cultured in OBM, OBM with FIT (bFGF, IGF-1, and TGF-β1), or OBM with SAG. FIT- and SAG-induced ALP activity significantly increased in patient-derived iPSCs compared with control iPSCs. (B) A sample of each supernatant was transferred to tubes for measurement of ALP activity as the hydrolysis of ρ-nitrophenyl phosphate using a LabAssay ALP kit and protein concentration using a Protein assay rapid kit. G-OF1iPS1 was showed a high ALP activity compared with 201B7,especially cultured in OBM with SAG. (C) KDiPSCs and G-OF1iPS1 cells were cultured in OBM (control), or OBM supplemented with 100 ng/ml BMP2/7 for 24 h. qRT-PCR analysis of RUNX2 was conducted using Premix Ex Taq reagent according to the manufacturer’s instructions. RUNX2 levels were normalized to those of GAPDH. A significant difference was observed in G-OF1iPS1. Especially, BMP2/7-induced RUNX2 expression promoted an approximate 2-fold increase in the number of G-OFiPSCs compared with KDiPSCs. Values represent the mean ± SD. Bonferroni correction for multiple comparisons was applied, * p

    Article Snippet: The stemness and pluripotency of the iPSCs were analyzed using reverse transcription polymerase chain reaction (RT-PCR) analysis with ExTaq DNA polymerase (Takara Biotechnology, Shiga, Japan).

    Techniques: Marker, ALP Assay, Derivative Assay, Cell Culture, Activity Assay, Protein Concentration, Quantitative RT-PCR, Expressing

    Distinct expression patterns of Hh-associated genes in control and patient-derived iPSCs under osteogenic conditions. We investigated two controls (KDiPS and 201B7) and four Gorlin iPS cases derived from different patients. Because we used single RT 2 Profiler PCR Array plate for each case, we did not performed statistical analysis. (A) Osteogeic induction promoted characteristic changes in the expression levels of the following genes associated with the canonical Hh pathway: GLI1 , GLI2 , GLI3 , and SHH . Compared with the basal conditions, GLI1 and GLI2 expression levels decreased in both controls (KDiPSCs and 201B7) and G-OFiPSCs grown in OBM, whereas GLI3 expression increased only in G-OFiPSCs grown in OBM. Basal expression levels of SHH was significantly lower in G-OFiPSCs compared with controls (KDiPSCs and 201B7). However, OBM enhanced SHH expression levels in G-OFiPSCs but not in control cells. (B) The basal expression levels of many Wnt genes were lower in patient-derived iPSCs than in control iPSCs. With the exception of WNT2 , all of the Wnt genes evaluated were upregulated in G-OFiPSCs grown in OBM and downregulated in control cells (KDiPSCs and 201B7) grown in OBM. (C) Changes in BMP2 , BMP6 , and RUNX2 expression levels were greater in patient-derived iPSCs than in controls (KD iPSCs and 201B7). The ΔΔC t value 10 days after OBM treatment (Post) was determined, and the fold-change relative to pre-induction (Pre) KDiPSCs was calculated for each gene.

    Journal: PLoS ONE

    Article Title: Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction

    doi: 10.1371/journal.pone.0186879

    Figure Lengend Snippet: Distinct expression patterns of Hh-associated genes in control and patient-derived iPSCs under osteogenic conditions. We investigated two controls (KDiPS and 201B7) and four Gorlin iPS cases derived from different patients. Because we used single RT 2 Profiler PCR Array plate for each case, we did not performed statistical analysis. (A) Osteogeic induction promoted characteristic changes in the expression levels of the following genes associated with the canonical Hh pathway: GLI1 , GLI2 , GLI3 , and SHH . Compared with the basal conditions, GLI1 and GLI2 expression levels decreased in both controls (KDiPSCs and 201B7) and G-OFiPSCs grown in OBM, whereas GLI3 expression increased only in G-OFiPSCs grown in OBM. Basal expression levels of SHH was significantly lower in G-OFiPSCs compared with controls (KDiPSCs and 201B7). However, OBM enhanced SHH expression levels in G-OFiPSCs but not in control cells. (B) The basal expression levels of many Wnt genes were lower in patient-derived iPSCs than in control iPSCs. With the exception of WNT2 , all of the Wnt genes evaluated were upregulated in G-OFiPSCs grown in OBM and downregulated in control cells (KDiPSCs and 201B7) grown in OBM. (C) Changes in BMP2 , BMP6 , and RUNX2 expression levels were greater in patient-derived iPSCs than in controls (KD iPSCs and 201B7). The ΔΔC t value 10 days after OBM treatment (Post) was determined, and the fold-change relative to pre-induction (Pre) KDiPSCs was calculated for each gene.

    Article Snippet: The stemness and pluripotency of the iPSCs were analyzed using reverse transcription polymerase chain reaction (RT-PCR) analysis with ExTaq DNA polymerase (Takara Biotechnology, Shiga, Japan).

    Techniques: Expressing, Derivative Assay, Polymerase Chain Reaction

    Generation of iPSCs from Gorlin syndrome fibroblasts (G-OFs). (A) We collected fibroblasts from surgically resected unaffected oral skin areas. Typical spindle shape fibroblasts were obtained (a). Fibroblasts (1 × 10 5 /well) were transfected with the Sendai virus vector SeVdp(KOSM)302L. The resulting iPSC colonies were selected 21–25 days after transfection and several clones were subsequently expanded (b). Stable clones (two clones of Patient 1, four clones of Patient 2, two clones of Patient 3, and six clones of Patient 4) exhibited the characteristic morphology of human iPSCs. Representative iPSCs (from passage number 6) are shown (c) (scale bar: 400 μm). (B) We confirmed the infection and silencing of SeVdp using RT-PCR. Cells expressed SeVdp mRNA one day after infection (TF). SeVdp was not detected in any G-OFs (F) or G-OFiPSCs. (C) RT-PCR analysis of embryonic stem cell marker genes in patient-derived iPSCs (G-OF1iPS clones 1 and 6; G-OF2iPS clones 3, 5, 8, and 10; G-OF3iPS clones 25 and 26; and G-OF4iPS clones 3, 8, 12, 15, 18, and 23) and control iPSCs (KDiPSCs). β-actin was used as a loading control. The target genes evaluated included OCT3/4 , SOX2 , NANOG , REX1 , c-MYC , and KLF4 .

    Journal: PLoS ONE

    Article Title: Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction

    doi: 10.1371/journal.pone.0186879

    Figure Lengend Snippet: Generation of iPSCs from Gorlin syndrome fibroblasts (G-OFs). (A) We collected fibroblasts from surgically resected unaffected oral skin areas. Typical spindle shape fibroblasts were obtained (a). Fibroblasts (1 × 10 5 /well) were transfected with the Sendai virus vector SeVdp(KOSM)302L. The resulting iPSC colonies were selected 21–25 days after transfection and several clones were subsequently expanded (b). Stable clones (two clones of Patient 1, four clones of Patient 2, two clones of Patient 3, and six clones of Patient 4) exhibited the characteristic morphology of human iPSCs. Representative iPSCs (from passage number 6) are shown (c) (scale bar: 400 μm). (B) We confirmed the infection and silencing of SeVdp using RT-PCR. Cells expressed SeVdp mRNA one day after infection (TF). SeVdp was not detected in any G-OFs (F) or G-OFiPSCs. (C) RT-PCR analysis of embryonic stem cell marker genes in patient-derived iPSCs (G-OF1iPS clones 1 and 6; G-OF2iPS clones 3, 5, 8, and 10; G-OF3iPS clones 25 and 26; and G-OF4iPS clones 3, 8, 12, 15, 18, and 23) and control iPSCs (KDiPSCs). β-actin was used as a loading control. The target genes evaluated included OCT3/4 , SOX2 , NANOG , REX1 , c-MYC , and KLF4 .

    Article Snippet: The stemness and pluripotency of the iPSCs were analyzed using reverse transcription polymerase chain reaction (RT-PCR) analysis with ExTaq DNA polymerase (Takara Biotechnology, Shiga, Japan).

    Techniques: Transfection, Plasmid Preparation, Clone Assay, Infection, Reverse Transcription Polymerase Chain Reaction, Marker, Derivative Assay

    Depletion of ChREBPβ attenuates glucose-stimulated β-cell proliferation in isolated rat islet cells. A and B : Isolated rat islet cells were treated with lipid-conjugated (Accell) siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h. Total RNA was collected and subjected to RT-PCR using primers specific for ChREBPβ or ChREBPα mRNAs. C : Protein extracts were subjected to immunoblotting using an antibody against ChREBPα and β-actin. D : To determine the effects of the siRNAs on ChREBP translocation, cells were treated with control or Accell siRNA against ChREBPβ cultured in 5.5 or 15 mmol/L glucose for 32 h and were fixed and stained for insulin, ChREBP, and DNA (DAPI). E : Isolated rat islet cells were treated with Accell siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h and fixed and stained with Ki67 and insulin. F : Quantification of the results in E , wherein at least 1,000 insulin-positive cells were counted. Results are from four different rat islet isolations. siChREBPβ-01 and -02, siRNAs directed against ChREBPβ; siCon, scramble control siRNA. Error bars are the SEM. * P

    Journal: Diabetes

    Article Title: Induction of the ChREBPβ Isoform Is Essential for Glucose-Stimulated β-Cell Proliferation

    doi: 10.2337/db15-0239

    Figure Lengend Snippet: Depletion of ChREBPβ attenuates glucose-stimulated β-cell proliferation in isolated rat islet cells. A and B : Isolated rat islet cells were treated with lipid-conjugated (Accell) siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h. Total RNA was collected and subjected to RT-PCR using primers specific for ChREBPβ or ChREBPα mRNAs. C : Protein extracts were subjected to immunoblotting using an antibody against ChREBPα and β-actin. D : To determine the effects of the siRNAs on ChREBP translocation, cells were treated with control or Accell siRNA against ChREBPβ cultured in 5.5 or 15 mmol/L glucose for 32 h and were fixed and stained for insulin, ChREBP, and DNA (DAPI). E : Isolated rat islet cells were treated with Accell siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h and fixed and stained with Ki67 and insulin. F : Quantification of the results in E , wherein at least 1,000 insulin-positive cells were counted. Results are from four different rat islet isolations. siChREBPβ-01 and -02, siRNAs directed against ChREBPβ; siCon, scramble control siRNA. Error bars are the SEM. * P

    Article Snippet: RNA Isolation and Quantitative RT-PCR Analysis RNA (0.5–1 μg) was used for reverse transcription, and RT-PCR analysis was conducted using The iTaq Universal SYBR Green Supermix (Bio-Rad, Cat. #172-5124) on the 7500 Applied Biosystems Real-Time System.

    Techniques: Isolation, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Translocation Assay, Staining

    Glucose induces ChREBPβ in primary rat and human islet cells. A – G : Dispersed rat islet cells were incubated in media containing 5.5 or 15 mmol/L glucose for 1 to 4 days, and the expression of the indicated genes were determined and shown as fold change relative to each day after normalizing to β-actin using the ΔΔCT method. H : Isolated human islet cells were cultured in media containing 5.5 or 15 mmol/L glucose for 1, 2, or 4 days. Total RNA was isolated and subjected to RT-PCR using primers specific for the indicated genes. The data are expressed as a fold change from 5.5 mmol/L glucose. I : Rat islets were isolated, dispersed, and incubated with lipid-conjugated Accell siRNA for 4 days. Total RNA was isolated and subjected to RT-PCR. Data are presented as relative to the scramble control (SiCon) 15 mmol/L treatment, after normalization to β-actin using the ΔΔCT method. Error bars are the SEM ( n = 3–4 for rat islets, n = 5–9 for human islets). ChREBPComm, ChREBP-common; siChREBPβ01 and -02, siRNAs directed against ChREBPβ. * P

    Journal: Diabetes

    Article Title: Induction of the ChREBPβ Isoform Is Essential for Glucose-Stimulated β-Cell Proliferation

    doi: 10.2337/db15-0239

    Figure Lengend Snippet: Glucose induces ChREBPβ in primary rat and human islet cells. A – G : Dispersed rat islet cells were incubated in media containing 5.5 or 15 mmol/L glucose for 1 to 4 days, and the expression of the indicated genes were determined and shown as fold change relative to each day after normalizing to β-actin using the ΔΔCT method. H : Isolated human islet cells were cultured in media containing 5.5 or 15 mmol/L glucose for 1, 2, or 4 days. Total RNA was isolated and subjected to RT-PCR using primers specific for the indicated genes. The data are expressed as a fold change from 5.5 mmol/L glucose. I : Rat islets were isolated, dispersed, and incubated with lipid-conjugated Accell siRNA for 4 days. Total RNA was isolated and subjected to RT-PCR. Data are presented as relative to the scramble control (SiCon) 15 mmol/L treatment, after normalization to β-actin using the ΔΔCT method. Error bars are the SEM ( n = 3–4 for rat islets, n = 5–9 for human islets). ChREBPComm, ChREBP-common; siChREBPβ01 and -02, siRNAs directed against ChREBPβ. * P

    Article Snippet: RNA Isolation and Quantitative RT-PCR Analysis RNA (0.5–1 μg) was used for reverse transcription, and RT-PCR analysis was conducted using The iTaq Universal SYBR Green Supermix (Bio-Rad, Cat. #172-5124) on the 7500 Applied Biosystems Real-Time System.

    Techniques: Incubation, Expressing, Isolation, Cell Culture, Reverse Transcription Polymerase Chain Reaction

    Map-based cloning of the z3 locus. a Physical mapping of the z3 locus. The z3 : Table S1. b Fine mapping of the z3 locus. The locus was further mapped within a 78-kb region between two markers, dCAPS7 and dCAPS2. Numbers below the line indicate F 2 recombinants at the marker regions. c Candidate genes (black boxes) in the 78-kb region. d T to C substitution of Z3 in the z3 mutant. Three exons and two introns are indicated as rectangles and lines, respectively. The position of the point mutation in the z3 mutant is at the third exon and represented by a black arrowhead. e Derived cleaved amplified polymorphic sequence (dCAPS) analysis of the point mutation in the z3 mutant. Aat ll was able to digest the genomic PCR products amplified from the WT, but not from the z3 mutant because of its T to C substitution in the Aat ll restriction enzyme site. M23, a mapping parent ‘Milyang23’; F1, F 1 hybrid ( z3 /Milyang23). f Z3 protein sequence. The predicted N-terminal signal peptide is marked with an open rectangle and the nine transmembrane domains (TMDs) are underlined. The yellow highlight represents regions predicted to be a CitMHS family. The position of the point mutation in the z3 mutant is highlighted in red

    Journal: Rice

    Article Title: The rice zebra3 (z3) mutation disrupts citrate distribution and produces transverse dark-green/green variegation in mature leaves

    doi: 10.1186/s12284-017-0196-8

    Figure Lengend Snippet: Map-based cloning of the z3 locus. a Physical mapping of the z3 locus. The z3 : Table S1. b Fine mapping of the z3 locus. The locus was further mapped within a 78-kb region between two markers, dCAPS7 and dCAPS2. Numbers below the line indicate F 2 recombinants at the marker regions. c Candidate genes (black boxes) in the 78-kb region. d T to C substitution of Z3 in the z3 mutant. Three exons and two introns are indicated as rectangles and lines, respectively. The position of the point mutation in the z3 mutant is at the third exon and represented by a black arrowhead. e Derived cleaved amplified polymorphic sequence (dCAPS) analysis of the point mutation in the z3 mutant. Aat ll was able to digest the genomic PCR products amplified from the WT, but not from the z3 mutant because of its T to C substitution in the Aat ll restriction enzyme site. M23, a mapping parent ‘Milyang23’; F1, F 1 hybrid ( z3 /Milyang23). f Z3 protein sequence. The predicted N-terminal signal peptide is marked with an open rectangle and the nine transmembrane domains (TMDs) are underlined. The yellow highlight represents regions predicted to be a CitMHS family. The position of the point mutation in the z3 mutant is highlighted in red

    Article Snippet: To determine the transcript levels of the Z3 gene by quantitative RT-PCR analysis, 20 μl of qRT-PCR mixture containing 2 μl of the first-strand cDNA mixture, 0.25 μM of the forward and reverse primers for each gene, and 10 μl of 2X QuantiTect LightCycler 480 SYBR Green I Master (Roche) were used. qRT-PCR analysis was performed on the Light Cycler 2.0 instrument (Roche Diagnostics, Germany).

    Techniques: Clone Assay, Marker, Mutagenesis, Derivative Assay, Amplification, Sequencing, Polymerase Chain Reaction

    MUC5AC gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by RT-PCR (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p

    Journal: Yonsei Medical Journal

    Article Title: Cholesterol Depletion in Cell Membranes of Human Airway Epithelial Cells Suppresses MUC5AC Gene Expression

    doi: 10.3349/ymj.2013.54.3.679

    Figure Lengend Snippet: MUC5AC gene expression after treatment with MβCD. NCI-H292 cells were pretreated with 1% MβCD for 1 hour, after which the media was freshly changed and cells were incubated with IL-1β (10 ng/mL) for 24 hours. MUC5AC gene expression was examined by RT-PCR (A) and real time-PCR (B) analysis. β 2 M served as the internal control. The expression of p-IL-1RI was examined by Western analysis (C). α-tubulin served as the internal control. IL-1β induces expression of MUC5AC , but this increased expression is significantly decreased by MβCD treatment (IL-1β+MβCD) ( p

    Article Snippet: Reverse transcription-polymerase chain reaction (RT-PCR) analysis of MUC5AC mRNA The total RNA was isolated from cells under each condition using TRI-reagent (Molecular Research Center, Cincinnati, OH, USA).

    Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot

    Effects of DIM and Herceptin on miR-200 family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA RT-PCR. ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P

    Journal: PLoS ONE

    Article Title: 3, 3?-diindolylmethane Enhances the Effectiveness of Herceptin against HER-2/Neu-Expressing Breast Cancer Cells

    doi: 10.1371/journal.pone.0054657

    Figure Lengend Snippet: Effects of DIM and Herceptin on miR-200 family. ( A ) Comparative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells by real-time miRNA RT-PCR. ( B and C ) Relative expression analysis of miR-200 in SKBR3 and MDA-MB-468 cells treated with 15 µM DIM, 0.75 µg/ml Herceptin or their combinations for 24 h. ( D ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on cell proliferation of SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as measured by MTT assay. ( E ) Efficacy of anti-miR-200s transfections as well the effect of DIM plus Herceptin (15 µM DIM and 0.75 µg/ml Herceptin for 72 hours) on expression levels of anti-miR-200s (miR-200a/miR-200b/miR-200c) was evaluated by real time RT-PCR. ( F ) Effect of anti-miR-200s (anti-miR-200a+anti-miR-200b+anti-miR-200c) on expression of FoxM1 and pAkt in SKBR3 cells treated with 15 µM DIM and 0.75 µg/ml Herceptin for 72 hours, as determined by western blotting. The numbers represent percent of corresponding control normalized to β-actin levels. Her, Herceptin; anti-miR-200s, anti-miR-200a+anti-miR-200b+anti-miR-200c. *, P

    Article Snippet: miRNA Real-time Reverse Transcription-PCR To verify the alterations in the expression of miR-200 in DIM and Herceptin-treated breast cancer cells, we performed RT-PCR analysis using TaqMan MicroRNA Assay Kit (Applied Biosystems) following the manufacturer’s protocol.

    Techniques: Expressing, Multiple Displacement Amplification, miRNA RT, Polymerase Chain Reaction, MTT Assay, Transfection, Quantitative RT-PCR, Western Blot

    NRAS is a target of miR-29a. A, Sequence of the miR-29a binding site within the human NRAS 3′-UTR and a schematic diagram of the reporter construct showing the entire NRAS 3′-UTR sequence and the mutant NRAS 3′-UTR sequence. The mutant nucleotides of the NRAS 3′-UTR are labeled in red. B, Luciferase assay on H1299 cells which were cotransfected with miR-NC or miR-29a and a luciferase reporter containing the full length of NRAS 3′-UTR (WT) or a mutant type (MT) harboring 4 mutant nucleotides of the miR-29a binding site. Luciferase activities were measured 24 hours posttransfection. MicroRNA-29a markedly suppressed luciferase activity in NRAS 3′-UTR wild type (WT) reporter constructs. C, The immunoblotting showed that expression levels of NRAS were decreased in cells with miR-29a overexpression. D, The expression of NRAS in adjacent normal tissues and human lung cancer specimens was determined by quantitative RT-PCR analysis, and fold changes were obtained from the ratio of NRAS to GAPDH levels. E, Spearman rank correlation analysis showed that the expression levels of NRAS and miR-29a in lung cancer tissues were inversely correlated. Data represent mean ± SD of three replicates. **Significant difference at P

    Journal: Technology in Cancer Research & Treatment

    Article Title: MicroRNA-29a Functions as a Tumor Suppressor and Increases Cisplatin Sensitivity by Targeting NRAS in Lung Cancer

    doi: 10.1177/1533033818758905

    Figure Lengend Snippet: NRAS is a target of miR-29a. A, Sequence of the miR-29a binding site within the human NRAS 3′-UTR and a schematic diagram of the reporter construct showing the entire NRAS 3′-UTR sequence and the mutant NRAS 3′-UTR sequence. The mutant nucleotides of the NRAS 3′-UTR are labeled in red. B, Luciferase assay on H1299 cells which were cotransfected with miR-NC or miR-29a and a luciferase reporter containing the full length of NRAS 3′-UTR (WT) or a mutant type (MT) harboring 4 mutant nucleotides of the miR-29a binding site. Luciferase activities were measured 24 hours posttransfection. MicroRNA-29a markedly suppressed luciferase activity in NRAS 3′-UTR wild type (WT) reporter constructs. C, The immunoblotting showed that expression levels of NRAS were decreased in cells with miR-29a overexpression. D, The expression of NRAS in adjacent normal tissues and human lung cancer specimens was determined by quantitative RT-PCR analysis, and fold changes were obtained from the ratio of NRAS to GAPDH levels. E, Spearman rank correlation analysis showed that the expression levels of NRAS and miR-29a in lung cancer tissues were inversely correlated. Data represent mean ± SD of three replicates. **Significant difference at P

    Article Snippet: Real-time reverse transcription polymerase chain reaction (RT-PCR) analysis for mature miR-29a was carried out using the RT Reagent Kit (Takara), according to the manufacturer’s instructions.

    Techniques: Sequencing, Binding Assay, Construct, Mutagenesis, Labeling, Luciferase, Activity Assay, Expressing, Over Expression, Quantitative RT-PCR

    Overexpression of miR-29a inhibits the ability of cell proliferation and colony formation in lung cancer cells. A and B, Relative expression levels of miR-29a in H1299/miR-29a, H1299/miR-NC, A549/miR-29a, and A549/miR-NC stable cell lines were confirmed by quantitative RT-PCR. C and D, Overexpression of miR-29a arrested cell proliferation in H1299 and A549 cells. E and F, MiR-29a overexpression reduced colony formation in H1299 and A549 cells. Data represent mean ± SD of 3 replicates. *Significant difference at P

    Journal: Technology in Cancer Research & Treatment

    Article Title: MicroRNA-29a Functions as a Tumor Suppressor and Increases Cisplatin Sensitivity by Targeting NRAS in Lung Cancer

    doi: 10.1177/1533033818758905

    Figure Lengend Snippet: Overexpression of miR-29a inhibits the ability of cell proliferation and colony formation in lung cancer cells. A and B, Relative expression levels of miR-29a in H1299/miR-29a, H1299/miR-NC, A549/miR-29a, and A549/miR-NC stable cell lines were confirmed by quantitative RT-PCR. C and D, Overexpression of miR-29a arrested cell proliferation in H1299 and A549 cells. E and F, MiR-29a overexpression reduced colony formation in H1299 and A549 cells. Data represent mean ± SD of 3 replicates. *Significant difference at P

    Article Snippet: Real-time reverse transcription polymerase chain reaction (RT-PCR) analysis for mature miR-29a was carried out using the RT Reagent Kit (Takara), according to the manufacturer’s instructions.

    Techniques: Over Expression, Expressing, Stable Transfection, Quantitative RT-PCR

    MicroRNA-29a is significantly downregulated in lung cancer tissues. A, Relative miR-29a expression levels were analyzed by quantitative RT-PCR in 38 pairs of human lung cancer tissues and adjacent normal tissues. U6 RNA level was used as an internal control. B, All samples were histologically classified by a clinical pathologist. Relative expression levels of miR-29a in different stages of cancer tissues. Data represent mean ± SD of 3 replicates. *Significant difference at P

    Journal: Technology in Cancer Research & Treatment

    Article Title: MicroRNA-29a Functions as a Tumor Suppressor and Increases Cisplatin Sensitivity by Targeting NRAS in Lung Cancer

    doi: 10.1177/1533033818758905

    Figure Lengend Snippet: MicroRNA-29a is significantly downregulated in lung cancer tissues. A, Relative miR-29a expression levels were analyzed by quantitative RT-PCR in 38 pairs of human lung cancer tissues and adjacent normal tissues. U6 RNA level was used as an internal control. B, All samples were histologically classified by a clinical pathologist. Relative expression levels of miR-29a in different stages of cancer tissues. Data represent mean ± SD of 3 replicates. *Significant difference at P

    Article Snippet: Real-time reverse transcription polymerase chain reaction (RT-PCR) analysis for mature miR-29a was carried out using the RT Reagent Kit (Takara), according to the manufacturer’s instructions.

    Techniques: Expressing, Quantitative RT-PCR

    Semiquantitative RT-PCR analysis of the expression of Bcl2 binding component 3/p53-up-regulated modulator of apoptosis ( Bbc3/PUMA ) in the presence of aluminum. Total RNA was analyzed by RT-PCR for the mRNA expressions of Bbc3/PUMA in primary cultured

    Journal: Environmental Health and Preventive Medicine

    Article Title: Gene expression in primary cultured astrocytes affected by aluminum: alteration of chaperons involved in protein folding

    doi: 10.1007/s12199-010-0161-2

    Figure Lengend Snippet: Semiquantitative RT-PCR analysis of the expression of Bcl2 binding component 3/p53-up-regulated modulator of apoptosis ( Bbc3/PUMA ) in the presence of aluminum. Total RNA was analyzed by RT-PCR for the mRNA expressions of Bbc3/PUMA in primary cultured

    Article Snippet: Validation of the effect of aluminum on the expression of selected genes was performed by reverse transcription (RT)-PCR analysis (BioRad) using apoptosis PCR bax/bcl2 multiplex primer sets (APO-PCR; Sigma–Aldrich, Saint Louis, MO).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Binding Assay, Cell Culture