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  • 99
    New England Biolabs luna universal quantitative pcr qpcr master mix
    Liver pathology showing significant nodular tumors in Slc39a8(+/neo) mice. A : wild-type (WT) mouse ( far left column ) was 20 mo of age; Slc39a8(+/neo) mice ( three right columns ) were 13, 18 and 21 mo of age, respectively. Macroscopic livers ( top row ) and microscopic hematoxylin-eosin (H E) staining ( bottom two rows ) is shown. Nodular tumors are visualized in parts or in the entire liver of Slc39a8(+/neo) mice. Out of 22 hypomorph mice, 10 were identified with similar nodular tumors. Scale bars: 500 μm ( middle row ); 100 μm ( bottom row ). B : hepatic Slc39a8 mRNA expression in WT ( n = 6) and Slc39a8(+/neo) mice ages 13–21 mo ( n = 7). C : typical immunohistochemistry (IHC) staining for α-fetoprotein (AFP) and proliferating cell nuclear antigen (PCNA) in WT and Slc39a8(+/neo) mice (20 mo of age) ( top ). Bottom : quantitative <t>PCR</t> analysis of Afp in WT and in nontumor portion and tumor of Slc39a8(+/neo) ( n = 3 for each of the three histograms). All data represent means ± SE, with 1.0 designated for WT mean expression. * P ≤ 0.05 by Student’s t -test. T, tumor.
    Luna Universal Quantitative Pcr Qpcr Master Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs luna one step universal quantitative pcr kit
    Plasma <t>lncRNA</t> TUG1 was upregulated in osteoporosis patients than in healthy participants. <t>RT-qPCR</t> results showed that plasma levels of lncRNA TUG1 were significantly higher in osteoporosis patients than in healthy participants (* p
    Luna One Step Universal Quantitative Pcr Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs luna universal probe qpcr master mix
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Probe Qpcr Master Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs luna universal probe qpcr kit
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Probe Qpcr Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs luna universal qpcr kit
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Qpcr Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad luna universal qpcr master mix kit
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Qpcr Master Mix Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam luna universal qpcr master mix
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Qpcr Master Mix, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega luna universal probe qpcr master mix
    Reduction in the <t>BoDV-1</t> load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by <t>RT-qPCR</t> analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P
    Luna Universal Probe Qpcr Master Mix, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Liver pathology showing significant nodular tumors in Slc39a8(+/neo) mice. A : wild-type (WT) mouse ( far left column ) was 20 mo of age; Slc39a8(+/neo) mice ( three right columns ) were 13, 18 and 21 mo of age, respectively. Macroscopic livers ( top row ) and microscopic hematoxylin-eosin (H E) staining ( bottom two rows ) is shown. Nodular tumors are visualized in parts or in the entire liver of Slc39a8(+/neo) mice. Out of 22 hypomorph mice, 10 were identified with similar nodular tumors. Scale bars: 500 μm ( middle row ); 100 μm ( bottom row ). B : hepatic Slc39a8 mRNA expression in WT ( n = 6) and Slc39a8(+/neo) mice ages 13–21 mo ( n = 7). C : typical immunohistochemistry (IHC) staining for α-fetoprotein (AFP) and proliferating cell nuclear antigen (PCNA) in WT and Slc39a8(+/neo) mice (20 mo of age) ( top ). Bottom : quantitative PCR analysis of Afp in WT and in nontumor portion and tumor of Slc39a8(+/neo) ( n = 3 for each of the three histograms). All data represent means ± SE, with 1.0 designated for WT mean expression. * P ≤ 0.05 by Student’s t -test. T, tumor.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Nutrient Sensing, Nutrition, and Metabolism: Hepatic ZIP8 deficiency is associated with disrupted selenium homeostasis, liver pathology, and tumor formation

    doi: 10.1152/ajpgi.00165.2018

    Figure Lengend Snippet: Liver pathology showing significant nodular tumors in Slc39a8(+/neo) mice. A : wild-type (WT) mouse ( far left column ) was 20 mo of age; Slc39a8(+/neo) mice ( three right columns ) were 13, 18 and 21 mo of age, respectively. Macroscopic livers ( top row ) and microscopic hematoxylin-eosin (H E) staining ( bottom two rows ) is shown. Nodular tumors are visualized in parts or in the entire liver of Slc39a8(+/neo) mice. Out of 22 hypomorph mice, 10 were identified with similar nodular tumors. Scale bars: 500 μm ( middle row ); 100 μm ( bottom row ). B : hepatic Slc39a8 mRNA expression in WT ( n = 6) and Slc39a8(+/neo) mice ages 13–21 mo ( n = 7). C : typical immunohistochemistry (IHC) staining for α-fetoprotein (AFP) and proliferating cell nuclear antigen (PCNA) in WT and Slc39a8(+/neo) mice (20 mo of age) ( top ). Bottom : quantitative PCR analysis of Afp in WT and in nontumor portion and tumor of Slc39a8(+/neo) ( n = 3 for each of the three histograms). All data represent means ± SE, with 1.0 designated for WT mean expression. * P ≤ 0.05 by Student’s t -test. T, tumor.

    Article Snippet: The quantitative (q)PCR for individual gene expression was carried out with Luna Universal qPCR Master Mix (New England Biolabs) and gene-specific primers; cycling conditions included 95°C for 1 min, followed by amplification for 40 cycles at 95°C for 15 s, and then 60°C for 30 s in the CFX96 Real-Time PCR system (Bio-Rad).

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

    Pcf11 Expression Is Regulated by IPA (A) Schematic of mouse Pcf11 gene. Multiple PASs are indicated. EPA, internal exonic polyadenylation; IPA, intronic polyadenylation; TPA, 3′ terminal exon polyadenylation. 3′READS data of brain, heart, and testis are shown. (B) Features of intron 1 compared with other introns in the mouse genome. Each feature has ten bins, and the bin containing intron 1 of Pcf11 is highlighted in red. Distance to TSS is based on all IPA sites in PolyA_DB. (C) RT-qPCR analysis of IPA and TPA isoform expression in siPcf11 versus siCtrl cells. Error bars are SD on the basis of three replicates. (D) Top: schematic showing two sgRNAs used to remove a region flanking the IPA site. Bottom: validation of IPA site knockout (IPA Pcf11 -KO) by genomic DNA PCR using primers flanking the KO region. WT, wild-type 4T1 cells. (E) RT-qPCR analysis of Pcf11 isoform expression in WT and IPA Pcf11 -KO cells. Error bars are SD on the basis of three replicates. (F) Western blot analysis of PCF11 in WT and IPA Pcf11 -KO cells. (G) 3′UTR APA changes in IPA Pcf11 -KO cells. (H) IPA changes in IPA Pcf11 -KO cells. IPA isoforms (all combined) were compared with TPA isoforms (all combined) in analysis. (I) IPA distribution maps of activated IPA events in IPA Pcf11 -KO cells. (J) Left: schematic of PASs around the transcription start site (TSS); right: metagene analysis of PAS usage around the TSS in WT and IPA Pcf1 1 -KO cells. (K) Gene expression changes in IPA Pcf11 -KO cells for different groups of genes on the basis of IPA site frequency in PolyA_DB. ***p

    Journal: Cell reports

    Article Title: Regulation of Intronic Polyadenylation by PCF11 Impacts mRNA Expression of Long Genes

    doi: 10.1016/j.celrep.2019.02.049

    Figure Lengend Snippet: Pcf11 Expression Is Regulated by IPA (A) Schematic of mouse Pcf11 gene. Multiple PASs are indicated. EPA, internal exonic polyadenylation; IPA, intronic polyadenylation; TPA, 3′ terminal exon polyadenylation. 3′READS data of brain, heart, and testis are shown. (B) Features of intron 1 compared with other introns in the mouse genome. Each feature has ten bins, and the bin containing intron 1 of Pcf11 is highlighted in red. Distance to TSS is based on all IPA sites in PolyA_DB. (C) RT-qPCR analysis of IPA and TPA isoform expression in siPcf11 versus siCtrl cells. Error bars are SD on the basis of three replicates. (D) Top: schematic showing two sgRNAs used to remove a region flanking the IPA site. Bottom: validation of IPA site knockout (IPA Pcf11 -KO) by genomic DNA PCR using primers flanking the KO region. WT, wild-type 4T1 cells. (E) RT-qPCR analysis of Pcf11 isoform expression in WT and IPA Pcf11 -KO cells. Error bars are SD on the basis of three replicates. (F) Western blot analysis of PCF11 in WT and IPA Pcf11 -KO cells. (G) 3′UTR APA changes in IPA Pcf11 -KO cells. (H) IPA changes in IPA Pcf11 -KO cells. IPA isoforms (all combined) were compared with TPA isoforms (all combined) in analysis. (I) IPA distribution maps of activated IPA events in IPA Pcf11 -KO cells. (J) Left: schematic of PASs around the transcription start site (TSS); right: metagene analysis of PAS usage around the TSS in WT and IPA Pcf1 1 -KO cells. (K) Gene expression changes in IPA Pcf11 -KO cells for different groups of genes on the basis of IPA site frequency in PolyA_DB. ***p

    Article Snippet: Real-time quantitative PCR (qPCR) Total RNA was extracted using TRIzol (Thermo Fisher) and treated with Turbo DNase (Thermo Fisher). cDNA was synthesized using oligo(dT) and M-MLV reverse transcriptase with 1.5–2 μg of RNA. cDNA was then mixed with real-time PCR primers and Luna qPCR master mix (NEB).

    Techniques: Expressing, Indirect Immunoperoxidase Assay, Quantitative RT-PCR, Knock-Out, Polymerase Chain Reaction, Western Blot

    Plasma lncRNA TUG1 was upregulated in osteoporosis patients than in healthy participants. RT-qPCR results showed that plasma levels of lncRNA TUG1 were significantly higher in osteoporosis patients than in healthy participants (* p

    Journal: Journal of Orthopaedic Surgery and Research

    Article Title: LncRNA TUG1 was upregulated in osteoporosis and regulates the proliferation and apoptosis of osteoclasts

    doi: 10.1186/s13018-019-1430-4

    Figure Lengend Snippet: Plasma lncRNA TUG1 was upregulated in osteoporosis patients than in healthy participants. RT-qPCR results showed that plasma levels of lncRNA TUG1 were significantly higher in osteoporosis patients than in healthy participants (* p

    Article Snippet: To detect the expression of lncRNA TUG1 and PTEN mRNA, Luna® Universal One-Step RT-qPCR Kit (NEB) was used to prepare PCR reaction systems.

    Techniques: Quantitative RT-PCR

    Histone methyltransferases EHMT1 and EHMT2 are upregulated in olaparib-resistant HGSOC. a Four olaparib resistant clones of PEO1-OR were analyzed by RNA-Seq. Of all enzymes involved in H3K9 methylation, EHMT1 (red squares), and KDM1B (green circles) were significantly changed in all four populations of PEO1-OR relative to PEO1. b – d RT-qPCR analysis of histone methyltransferases EHMT1 and EHMT2 , and zinc-finger gene ZNF644 in PEO1 and PEO1-OR cells (mean ± SD, n = 3, unpaired t test). e Protein lysates from PEO1 and PEO1-OR cells were analyzed by immunoblot for EHMT1, EHMT2, and Actin loading control. f , g Patient-derived HGSOC ascites cells were injected intraperitoneally into NSG mice. After 21-day treatment with olaparib or vehicle control, mice were sacrificed and ascites cells were collected and analyzed by RT-qPCR for EHMT1 and EHMT2 (mRNA expression is normalized to GAPDH and plotted as mean ± SD, n = 3 technical PCR replicates, unpaired test; numbers below bars are mouse ear-tag numbers). h Protein lysates from ascites cells were analyzed by immunoblot for EHMT1, EHMT2, and Actin loading control (ear tag numbers correspond with mRNA data)

    Journal: Clinical Epigenetics

    Article Title: Histone methyltransferases EHMT1 and EHMT2 (GLP/G9A) maintain PARP inhibitor resistance in high-grade serous ovarian carcinoma

    doi: 10.1186/s13148-019-0758-2

    Figure Lengend Snippet: Histone methyltransferases EHMT1 and EHMT2 are upregulated in olaparib-resistant HGSOC. a Four olaparib resistant clones of PEO1-OR were analyzed by RNA-Seq. Of all enzymes involved in H3K9 methylation, EHMT1 (red squares), and KDM1B (green circles) were significantly changed in all four populations of PEO1-OR relative to PEO1. b – d RT-qPCR analysis of histone methyltransferases EHMT1 and EHMT2 , and zinc-finger gene ZNF644 in PEO1 and PEO1-OR cells (mean ± SD, n = 3, unpaired t test). e Protein lysates from PEO1 and PEO1-OR cells were analyzed by immunoblot for EHMT1, EHMT2, and Actin loading control. f , g Patient-derived HGSOC ascites cells were injected intraperitoneally into NSG mice. After 21-day treatment with olaparib or vehicle control, mice were sacrificed and ascites cells were collected and analyzed by RT-qPCR for EHMT1 and EHMT2 (mRNA expression is normalized to GAPDH and plotted as mean ± SD, n = 3 technical PCR replicates, unpaired test; numbers below bars are mouse ear-tag numbers). h Protein lysates from ascites cells were analyzed by immunoblot for EHMT1, EHMT2, and Actin loading control (ear tag numbers correspond with mRNA data)

    Article Snippet: Reverse-transcriptase quantitative PCR RNA was isolated from cells using the RNeasy Plus Mini Kit (Qiagen). mRNA expression was determined using SYBR green Luna One Step reverse-transcriptase quantitative PCR (RT-qPCR) Kit (New England BioLabs) on a C1000 Touch (Bio-Rad) or QuantStudio 6 (Applied Biosystems) thermocycler.

    Techniques: Clone Assay, RNA Sequencing Assay, Methylation, Quantitative RT-PCR, Derivative Assay, Injection, Mouse Assay, Expressing, Polymerase Chain Reaction

    Expression level of differentially expressed lncRNAs in NSTEMI and STEMI patients. ENST00000508020.2, LNC_002011, LNC_000303 LNC_000898, ENST00000573866.2, and ENST00000562710.1 was detected by qPCR and normalized by GAPHD expression in STEMI patients and NSTEMI patients (All P values were less than .05). lncRNAs = long noncoding RNAs, NSTEMI = non-ST-elevation myocardial infarction, STEMI = ST-elevation myocardial infarction.

    Journal: Medicine

    Article Title: Differential expression of circulating long non-coding RNAs in patients with acute myocardial infarction

    doi: 10.1097/MD.0000000000013066

    Figure Lengend Snippet: Expression level of differentially expressed lncRNAs in NSTEMI and STEMI patients. ENST00000508020.2, LNC_002011, LNC_000303 LNC_000898, ENST00000573866.2, and ENST00000562710.1 was detected by qPCR and normalized by GAPHD expression in STEMI patients and NSTEMI patients (All P values were less than .05). lncRNAs = long noncoding RNAs, NSTEMI = non-ST-elevation myocardial infarction, STEMI = ST-elevation myocardial infarction.

    Article Snippet: 2.6 Validation by real-time quantitative PCR cDNA was synthesized and Real-Time Quantitative PCR (RT-qPCR) of lncRNAs expression level was performed using Luna Universal One-Step RT-qPCR kits (New England Biolabs, MA) according to the manufacturer's protocol.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Reduction in the BoDV-1 load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P

    Journal: Frontiers in Microbiology

    Article Title: A Small Interfering RNA Cocktail Targeting the Nucleoprotein and Large Protein Genes Suppresses Borna Disease Virus Infection

    doi: 10.3389/fmicb.2019.02781

    Figure Lengend Snippet: Reduction in the BoDV-1 load by siRNAs targeting N and M/G/L mRNAs. (A,B) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 mRNA amounts. 293T/BoDV cells were treated with the indicated siRNA for 2 days. The amounts of N (A) and M/G/L (B) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (C) Effects of siRNAs targeting BoDV-1 mRNAs on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (D,E) Effects of the siRNA cocktail (TD-Borna) targeting BoDV-1 N and M/G/L mRNAs on BoDV-1 mRNAs. 293T/BoDV cells were treated with TD-Borna for 2 days. The amounts of N (D) and M/G/L (E) mRNAs in 293T/BoDV cells were determined by RT-qPCR analyses. (F) Effects of TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (G) Protocol for long-term TD-Borna treatment in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0, 6, and 12. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) every 3 days. (H) Effects of long-term TD-Borna treatment on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗∗ P

    Article Snippet: RT-qPCR assays for BoDV-1 L mRNA and BoDV-1 gRNA were carried out using Luna Universal Probe qPCR Master Mix (New England Biolabs, Ipswich, United States) and the BoDV-1-specific primers and probe.

    Techniques: Quantitative RT-PCR, Transfection

    Reduction in the BoDV-1 load by the combined use of T-705 and TD-Borna. (A) Protocol for long-term treatment using T-705 and TD-Borna in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0 and 6. On Day 0, T-705 was added at 1 h after the transfection. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) with or without T-705 on Days 2, 5, and 8. (B) Effects of long-term treatment with T-705 and TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (C) The amount of the N protein in 293T/BoDV cells treated with T-705 and TD-Borna. 293T/BoDV cells were treated as indicated in panel (A) . After 5 days of treatment, the amount of the N protein was determined by western blotting using anti-N and anti-tubulin antibodies. (D) Quantification of the amount of the N protein in panel (C) . The band intensity of the N protein in each sample was normalized with that of tubulin. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗ P

    Journal: Frontiers in Microbiology

    Article Title: A Small Interfering RNA Cocktail Targeting the Nucleoprotein and Large Protein Genes Suppresses Borna Disease Virus Infection

    doi: 10.3389/fmicb.2019.02781

    Figure Lengend Snippet: Reduction in the BoDV-1 load by the combined use of T-705 and TD-Borna. (A) Protocol for long-term treatment using T-705 and TD-Borna in 293T/BoDV cells. 293T/BoDV cells were seeded in 12-well plates (2 × 10 5 cells/well) on Day –1. TD-Borna was transfected on Days 0 and 6. On Day 0, T-705 was added at 1 h after the transfection. The cells were passaged into new 12-well plates (2 × 10 5 cells/well) with or without T-705 on Days 2, 5, and 8. (B) Effects of long-term treatment with T-705 and TD-Borna on BoDV-1 replication. The amount of BoDV-1 gRNA in 293T/BoDV cells was determined by RT-qPCR analyses. (C) The amount of the N protein in 293T/BoDV cells treated with T-705 and TD-Borna. 293T/BoDV cells were treated as indicated in panel (A) . After 5 days of treatment, the amount of the N protein was determined by western blotting using anti-N and anti-tubulin antibodies. (D) Quantification of the amount of the N protein in panel (C) . The band intensity of the N protein in each sample was normalized with that of tubulin. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. ∗ P

    Article Snippet: RT-qPCR assays for BoDV-1 L mRNA and BoDV-1 gRNA were carried out using Luna Universal Probe qPCR Master Mix (New England Biolabs, Ipswich, United States) and the BoDV-1-specific primers and probe.

    Techniques: Transfection, Quantitative RT-PCR, Western Blot

    Reduction in the BoDV-1 load by TD-Borna in neuronal and non-neuronal cell lines. (A,B) Effect of T-705 on BoDV-1 replication in 293T/BoDV (A) and SH-SY5Y/BoDV (B) cells. (C,D) Effect of TD-Borna on BoDV-1 replication in 293T/BoDV (C) and SH-SY5Y/BoDV (D) cells. The cells were treated with T-705 (A,B) or TD-Borna (C,D) for 2 days. The amount of BoDV-1 gRNA in the cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. **** P

    Journal: Frontiers in Microbiology

    Article Title: A Small Interfering RNA Cocktail Targeting the Nucleoprotein and Large Protein Genes Suppresses Borna Disease Virus Infection

    doi: 10.3389/fmicb.2019.02781

    Figure Lengend Snippet: Reduction in the BoDV-1 load by TD-Borna in neuronal and non-neuronal cell lines. (A,B) Effect of T-705 on BoDV-1 replication in 293T/BoDV (A) and SH-SY5Y/BoDV (B) cells. (C,D) Effect of TD-Borna on BoDV-1 replication in 293T/BoDV (C) and SH-SY5Y/BoDV (D) cells. The cells were treated with T-705 (A,B) or TD-Borna (C,D) for 2 days. The amount of BoDV-1 gRNA in the cells was determined by RT-qPCR analyses. Mock, the scrambled siRNA-treated control. Values are expressed as the mean ± SE of three independent experiments. **** P

    Article Snippet: RT-qPCR assays for BoDV-1 L mRNA and BoDV-1 gRNA were carried out using Luna Universal Probe qPCR Master Mix (New England Biolabs, Ipswich, United States) and the BoDV-1-specific primers and probe.

    Techniques: Quantitative RT-PCR