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  • 99
    Thermo Fisher trizol reagent
    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the <t>TRIzol</t> reagent (Invitrogen) to extract total <t>RNA</t> and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P
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    TaKaRa primescript rt reagent kit
    QODG insignificantly regulates the VEGF-triggered activation of VEGFR1 and VEGFR2 mRNAs expression in HUVECs. HUVECs (5×10 5 cells/well) were treated with or without VEGF (50 ng/mL) and DMSO (0.1%) or various concentrations of QODG (20, 60, 180 µM) for 24 h. RNA was extracted with TRIzol® Reagent (Invitrogen, Life Technologies, Grand Island, NY, USA), reverse transcribed with <t>PrimeScript™</t> RT reagent kit (TaKaRa, Otsu, Shiga, Japan), and quantitated by qRT-PCR using SYBR® Premix Ex Taq™ (TaKaRa, Otsu, Shiga, Japan). Cells receiving only DMSO (0.1%) served as a vehicle control. The levels of VEGFR1 and VEGFR2 mRNAs are normalized by β-actin and expressed as percentages of the vehicle control (100%) in mean ± SD from three independent experiments in triplicates. P > 0.05 vs. VEGF-treated control.
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    TaKaRa sybr premix ex taq
    qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a <t>SYBR</t> Premix Ex <t>Taq</t> kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.
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    TaKaRa primescript rt master mix
    qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a <t>SYBR</t> Premix Ex <t>Taq</t> kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.
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    97
    Bio-Rad qrt pcr
    qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a <t>SYBR</t> Premix Ex <t>Taq</t> kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.
    Qrt Pcr, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 6815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa gdna eraser
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
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    Thermo Fisher 7500 real time pcr system
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
    7500 Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 61883 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi 7500 real time pcr system
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
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    Thermo Fisher steponeplus real time pcr system
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
    Steponeplus Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad cfx96 real time pcr detection system
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
    Cfx96 Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 27018 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa mir x mirna first strand synthesis kit
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
    Mir X Mirna First Strand Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1980 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher revertaid first strand cdna synthesis kit
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
    Revertaid First Strand Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 40122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy mini kit
    <t>DNA</t> damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.
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    Image Search Results


    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Journal: International Journal of Biological Sciences

    Article Title: ZIP4, a Novel Determinant of Tumor Invasion in Hepatocellular Carcinoma, Contributes to Tumor Recurrence after Liver Transplantation

    doi: 10.7150/ijbs.7401

    Figure Lengend Snippet: (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Article Snippet: Real-time polymerase chain reaction RNA was extracted using the TRIzol reagent (Invitrogen) and used for cDNA synthesis with a M-MLV Reverse Transcriptase (TaKaRa, Dalian, China).

    Techniques: Microscopy, Incubation, Cell Culture, Staining, Negative Control, shRNA, Over Expression

    Co-segregation of the T-DNA with the chlorina phenotype. (A) Schematic representation of the T-DNA insertion upstream of OsSHMT1 (exons = black/red boxes; introns = black lines) and the domain organization of OsSHMT1 ( http://www.uniprot.org/uniprot/Q6EPF0 ). ATG, start codon; TAG, termination codon; RB, right border; LB, left border. P1, P2, and P3 are the primers used for genotyping. (B ) Co-segregation analysis of the genotype and phenotype in a segregating population. All plants homozygous for the T-DNA insertion showed a chlorotic phenotype, indicating that the recessive mutation was caused by T-DNA insertion. Numbers represent the different plants tested. P1/P2, PCR using primers P1 and P2; P1/P3, PCR using primers P1 and P3; He, hemizygous; Ho, homozygous; W, wild type. “+” represents the wild-type phenotype; “−” represents the mutant phenotype. Plants 2, 5, 9, and 13 were homozygous because only the 0.9-kb band was amplified using primers P1 and P2. Plants 4, 7, 11, 15, and 18 were designated as wild type because only the 0.7-kb band was amplified using primers P1 and P3. In contrast, both the 0.7- and 0.9-kb bands were amplified from plants 1, 3, 6, 8, 10, 12, 14, 16, 17, and 19, indicating that they were heterozygotic. (C) Analyses of OsSHMT1 expression in wild-type and oscdm1 mutant plants using actin as a control. The RT-PCR primers P4 and P5 for OsSHMT1 span introns 7 and 8. RT-PCR using RNA from homozygous oscdm1 plants did not detect full-length OsSHMT1 transcripts.

    Journal: Frontiers in Genetics

    Article Title: The molecular cloning and clarification of a photorespiratory mutant, oscdm1, using enhancer trapping

    doi: 10.3389/fgene.2015.00226

    Figure Lengend Snippet: Co-segregation of the T-DNA with the chlorina phenotype. (A) Schematic representation of the T-DNA insertion upstream of OsSHMT1 (exons = black/red boxes; introns = black lines) and the domain organization of OsSHMT1 ( http://www.uniprot.org/uniprot/Q6EPF0 ). ATG, start codon; TAG, termination codon; RB, right border; LB, left border. P1, P2, and P3 are the primers used for genotyping. (B ) Co-segregation analysis of the genotype and phenotype in a segregating population. All plants homozygous for the T-DNA insertion showed a chlorotic phenotype, indicating that the recessive mutation was caused by T-DNA insertion. Numbers represent the different plants tested. P1/P2, PCR using primers P1 and P2; P1/P3, PCR using primers P1 and P3; He, hemizygous; Ho, homozygous; W, wild type. “+” represents the wild-type phenotype; “−” represents the mutant phenotype. Plants 2, 5, 9, and 13 were homozygous because only the 0.9-kb band was amplified using primers P1 and P2. Plants 4, 7, 11, 15, and 18 were designated as wild type because only the 0.7-kb band was amplified using primers P1 and P3. In contrast, both the 0.7- and 0.9-kb bands were amplified from plants 1, 3, 6, 8, 10, 12, 14, 16, 17, and 19, indicating that they were heterozygotic. (C) Analyses of OsSHMT1 expression in wild-type and oscdm1 mutant plants using actin as a control. The RT-PCR primers P4 and P5 for OsSHMT1 span introns 7 and 8. RT-PCR using RNA from homozygous oscdm1 plants did not detect full-length OsSHMT1 transcripts.

    Article Snippet: Reverse transcription (RT)-PCR and quantitative real-time RT-PCR (qRT-PCR) To study the OsSHMT1 expression level, total RNA was extracted using TRIzol reagent (Invitrogen) from the leaves of wild-type and oscdm1 plants.

    Techniques: Mutagenesis, Polymerase Chain Reaction, Amplification, Expressing, Reverse Transcription Polymerase Chain Reaction

    QODG insignificantly regulates the VEGF-triggered activation of VEGFR1 and VEGFR2 mRNAs expression in HUVECs. HUVECs (5×10 5 cells/well) were treated with or without VEGF (50 ng/mL) and DMSO (0.1%) or various concentrations of QODG (20, 60, 180 µM) for 24 h. RNA was extracted with TRIzol® Reagent (Invitrogen, Life Technologies, Grand Island, NY, USA), reverse transcribed with PrimeScript™ RT reagent kit (TaKaRa, Otsu, Shiga, Japan), and quantitated by qRT-PCR using SYBR® Premix Ex Taq™ (TaKaRa, Otsu, Shiga, Japan). Cells receiving only DMSO (0.1%) served as a vehicle control. The levels of VEGFR1 and VEGFR2 mRNAs are normalized by β-actin and expressed as percentages of the vehicle control (100%) in mean ± SD from three independent experiments in triplicates. P > 0.05 vs. VEGF-treated control.

    Journal: PLoS ONE

    Article Title: Quercetin-4?-O-?-D-glucopyranoside (QODG) Inhibits Angiogenesis by Suppressing VEGFR2-Mediated Signaling in Zebrafish and Endothelial Cells

    doi: 10.1371/journal.pone.0031708

    Figure Lengend Snippet: QODG insignificantly regulates the VEGF-triggered activation of VEGFR1 and VEGFR2 mRNAs expression in HUVECs. HUVECs (5×10 5 cells/well) were treated with or without VEGF (50 ng/mL) and DMSO (0.1%) or various concentrations of QODG (20, 60, 180 µM) for 24 h. RNA was extracted with TRIzol® Reagent (Invitrogen, Life Technologies, Grand Island, NY, USA), reverse transcribed with PrimeScript™ RT reagent kit (TaKaRa, Otsu, Shiga, Japan), and quantitated by qRT-PCR using SYBR® Premix Ex Taq™ (TaKaRa, Otsu, Shiga, Japan). Cells receiving only DMSO (0.1%) served as a vehicle control. The levels of VEGFR1 and VEGFR2 mRNAs are normalized by β-actin and expressed as percentages of the vehicle control (100%) in mean ± SD from three independent experiments in triplicates. P > 0.05 vs. VEGF-treated control.

    Article Snippet: RNA samples were quantified at OD260/280 , and RNA was introduced to reverse transcribe to single-stranded cDNA using PrimeScript™ RT reagent kit (TaKaRa, Otsu, Shiga, Japan), followed by qTR-PCR using the SYBR® Premix Ex Taq™ (TaKaRa, Otsu, Shiga, Japan) in Mastercycler® ep gradient realplex2 Real-Time PCR System (Eppendorf, Hamburg, Germany).

    Techniques: Activation Assay, Expressing, Quantitative RT-PCR

    qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a SYBR Premix Ex Taq kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.

    Journal: Scientific Reports

    Article Title: Overexpression of the Eggplant (Solanum melongena) NAC Family Transcription Factor SmNAC Suppresses Resistance to Bacterial Wilt

    doi: 10.1038/srep31568

    Figure Lengend Snippet: qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a SYBR Premix Ex Taq kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.

    Article Snippet: Quantitative reverse-transcription (qRT-PCR) analysis Quantitative reverse-transcription PCR (qPCR) was performed using gene-specific primers ( ) and a SYBR Premix Ex Taq kit (TaKaRa, Dalian, China), following the manufacturer’s protocols.

    Techniques: Quantitative RT-PCR, Expressing, Transgenic Assay, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    DNA damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.

    Journal: Scientific Reports

    Article Title: Arabidopsis PARG1 is the key factor promoting cell survival among the enzymes regulating post-translational poly(ADP-ribosyl)ation

    doi: 10.1038/srep15892

    Figure Lengend Snippet: DNA damage level is higher in the parg1-4 mutant than that in Col-0 plants under genotoxic stress. ( A ) Expression level changes of genes involved in DNA double-strand break repair in roots of Col-0 and parg1-4 seedlings. The data represent the mean values of three replicates ± SD. ( C ) control; ( B ) bleomycin at 20 μg ml −1 . ( B ) Comet assay of the DNA damage levels of Col-0 and parg1-4 seedlings grown on plates with or without (control) 20 μg ml −1 bleomycin for 10 days. The percentage of DNA in comet tails was analyzed and quantified by CASP software ( http://sourceforge.net/projects/casp/ ) and used as an indicator of DNA damage level. 100 nuclei for each treatment were randomly selected and imaged. The bar size represents proportion of nuclei falling into the ranges of damage level indicated by different colors, and the images with percentages beside it indicate the examples of damaged nuclei. ( C ) DNA fragmentation assay showed that genomic DNA is more damaged in parg1-4 than in Col-0.

    Article Snippet: Complementary DNA (cDNA) was synthesized using PrimeScript RT-PCR Kit with gDNA eraser (Takara, Japan), and then used for qRT-PCR.

    Techniques: Mutagenesis, Expressing, Single Cell Gel Electrophoresis, Software, DNA Fragmentation Assay