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  • 95
    Millipore cas9 mrnas
    RGEN-induced Prkdc gene disruption in mice. ( A ) Representative T7E1 assays demonstrating mutagenesis efficiencies of <t>Cas9</t> <t>mRNA</t> plus Prkdc -specific sgRNA that were delivered via intracytoplasmic injection into one-cell-stage mouse embryos. Numbers indicate
    Cas9 Mrnas, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Shanghai Generay Biotech mrnas
    RGEN-induced Prkdc gene disruption in mice. ( A ) Representative T7E1 assays demonstrating mutagenesis efficiencies of <t>Cas9</t> <t>mRNA</t> plus Prkdc -specific sgRNA that were delivered via intracytoplasmic injection into one-cell-stage mouse embryos. Numbers indicate
    Mrnas, supplied by Shanghai Generay Biotech, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa mrnas
    Effects of chitosan nanofibers on the induction of osteoblast differentiation-related OPN, OCN, and ALP gene expression. Notes: Human osteoblast-like MG63 cells were seeded on chitosan nanofiber scaffolds for 1, 3, and 5 days. ( A ) Analyses of OPN, OCN, and ALP <t>mRNAs</t> were conducted using <t>RT-PCR;</t> β-actin mRNA was analyzed as an internal control. The DNA bands were quantified and statistically analyzed ( B – D ). Each value represents the mean ± standard error of the mean from four independent experiments. * indicates that values significantly differed from the respective control, P
    Mrnas, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 2412 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher mrnas
    Host cell mRNA synthesis and translation are unaffected by compound JMN3-003. A ) Relative TaqMan RT-PCR-based quantitation of three unstable cellular <t>mRNAs</t> (MCL1, <t>ASB7,</t> MKP-1) after exposure of cells to JMN3-003 for six hours. Controls were treated with Actinomycine D (Act D) for comparison. C T values are expressed relative to vehicle-treated samples and reflect averages of three independent experiments, each analyzed in triplicate, ± SD. B–D ) Expression of virus-encoded but not host cell or plasmid-encoded viral proteins is blocked by JMN3-003. Immunodetection of transiently expressed MeV-F ( B ), virus-encoded MeV-F ( C ), and virus-encoded influenza A/WSN M2 ( D ) in cell lysates after incubation of cells in the presence of compound or vehicle only (DMSO) for 30 hours. As internal cellular standard, membranes were probed for GAPDH in parallel. Numbers correspond to average densitometric quantitations ± SD of three experiments, representative immunoblots are shown. (ND: not determined).
    Mrnas, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 18500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Agilent technologies mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 906 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cell Matrix Inc mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by Cell Matrix Inc, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Horizon Discovery mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 92/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 694 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pacific Biosciences mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by Pacific Biosciences, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    SAS institute mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by SAS institute, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    ValueGene mrnas
    Expression profiles and enrichment Analysis of <t>mRNAs/lncRNAs</t> regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p
    Mrnas, supplied by ValueGene, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    VANGL2 LTD mrnas
    Maternal <t>Vangl2</t> and aPKC depletion disrupts the pattern of expression of specific zygotic genes. ( A ) Relative expression levels of Siamois , Xnr3 and Foxi1e <t>mRNAs</t> in control (Uninj) and Vangl2-depleted (VG–) embryos at the late blastula (st9) and
    Mrnas, supplied by VANGL2 LTD, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Bio-Rad mrnas
    Expression of HSI2 , HSL1 , and seed maturation genes in Col-0 and KK seedlings. Total RNAs were isolated from 4-, 5-, 7-, and 9-day-old Col-0 and KK seedlings grown on medium containing 1% sucrose. The levels of various <t>mRNAs</t> were determined by quantitative real-time <t>RT-PCR</t> using ACTIN2 mRNA as an internal reference. Results represent the average from two independent isolations of RNA ± SD.
    Mrnas, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1617 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Stemgent mrnas
    Schematic representation of iPSC derivation protocol and the generation of iPSCs from HSF and HUVECs. (A) On day 0, the parent cells, HSF and HVECs cells were treated separately with Stemcircle, a plasmid containing <t>Oct4,</t> Nanog, Sox2 and Lin28 for 24 hours and subsequently, a cocktail of <t>mRNAs</t> containing Oct4, Sox2, Klf4, cMyc, Lin28 for up to day 12. The colonies were allowed to expand in between day 13 and 16. On day 17, the fully expanded colonies were picked and grown. HSF-Human skin fibroblast, HUVECs-Human umbilical vein endothelial cells, D-day. (B1) Phage contrast microscopic image of iPSC-derived from HSF (hf-iPSCs). (B2) Live staining of Tra1-60+ cells from hf-iPSCs. (C1) Phage contrast microscopic image of iPSC-derived from HUVECs (he-iPSCs). (C2) Live staining of Tra1-60+ cells from he-iPSCs. (D) Transfection efficiency of iPSCs generated from HSF by flow cytometry analysis at passage 0 (P0). The data show 4% of cells were positive for either Oct4 or Sox2. (E) Quantification of Tra1-60+ cells by flow cytometry at P9. The data show 73.2% of cells are Tra1-60+. Data shown are representative of three independent experiments, **p
    Mrnas, supplied by Stemgent, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Sangon Biotech mrnas
    Schematic representation of iPSC derivation protocol and the generation of iPSCs from HSF and HUVECs. (A) On day 0, the parent cells, HSF and HVECs cells were treated separately with Stemcircle, a plasmid containing <t>Oct4,</t> Nanog, Sox2 and Lin28 for 24 hours and subsequently, a cocktail of <t>mRNAs</t> containing Oct4, Sox2, Klf4, cMyc, Lin28 for up to day 12. The colonies were allowed to expand in between day 13 and 16. On day 17, the fully expanded colonies were picked and grown. HSF-Human skin fibroblast, HUVECs-Human umbilical vein endothelial cells, D-day. (B1) Phage contrast microscopic image of iPSC-derived from HSF (hf-iPSCs). (B2) Live staining of Tra1-60+ cells from hf-iPSCs. (C1) Phage contrast microscopic image of iPSC-derived from HUVECs (he-iPSCs). (C2) Live staining of Tra1-60+ cells from he-iPSCs. (D) Transfection efficiency of iPSCs generated from HSF by flow cytometry analysis at passage 0 (P0). The data show 4% of cells were positive for either Oct4 or Sox2. (E) Quantification of Tra1-60+ cells by flow cytometry at P9. The data show 73.2% of cells are Tra1-60+. Data shown are representative of three independent experiments, **p
    Mrnas, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 91/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Stratagene mrnas
    miR-483-5p induces promoter-specific <t>IGF2</t> transcription. ( A ) Scatter plot (log) of mRNA profiles of MHH-ES-1 cells transfected with miR-483-5p ( Y -axis) versus scrambled control ( X -axis). Transcripts expressed in both samples (red), one sample (blue), or neither sample (yellow) are shown (as determined by default settings of the Affymetrix GCOS software), with green lines indicating twofold, threefold, 10-fold, and 30-fold differential expression between the two samples. The arrow points to signals from three independent IGF2 <t>mRNAs</t> probes. ( B ) Genes that are changed (either up-regulated or down-regulated) more than twofold in MHH-ES-1 cells transfected with a miR-483-5p mimic compared with scrambled control. ( C ) Time course of IGF2 mRNA expression after transfection of MHH-ES-1 with a miR-483-5p mimic, as measured by qRT–PCR. Expression is relative to levels at day 0. ( D ) Induction of IGF2 mRNA by miR-483-5p in two human sarcoma cell lines (MMH-ES-1 and RD) and two mouse sarcoma lines (F4328 and F4864), as measured by qRT–PCR. The asterisk denotes a significant difference ( P
    Mrnas, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 512 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Waters Corporation mrnas
    miR-483-5p induces promoter-specific <t>IGF2</t> transcription. ( A ) Scatter plot (log) of mRNA profiles of MHH-ES-1 cells transfected with miR-483-5p ( Y -axis) versus scrambled control ( X -axis). Transcripts expressed in both samples (red), one sample (blue), or neither sample (yellow) are shown (as determined by default settings of the Affymetrix GCOS software), with green lines indicating twofold, threefold, 10-fold, and 30-fold differential expression between the two samples. The arrow points to signals from three independent IGF2 <t>mRNAs</t> probes. ( B ) Genes that are changed (either up-regulated or down-regulated) more than twofold in MHH-ES-1 cells transfected with a miR-483-5p mimic compared with scrambled control. ( C ) Time course of IGF2 mRNA expression after transfection of MHH-ES-1 with a miR-483-5p mimic, as measured by qRT–PCR. Expression is relative to levels at day 0. ( D ) Induction of IGF2 mRNA by miR-483-5p in two human sarcoma cell lines (MMH-ES-1 and RD) and two mouse sarcoma lines (F4328 and F4864), as measured by qRT–PCR. The asterisk denotes a significant difference ( P
    Mrnas, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Unigene mrnas
    miR-483-5p induces promoter-specific <t>IGF2</t> transcription. ( A ) Scatter plot (log) of mRNA profiles of MHH-ES-1 cells transfected with miR-483-5p ( Y -axis) versus scrambled control ( X -axis). Transcripts expressed in both samples (red), one sample (blue), or neither sample (yellow) are shown (as determined by default settings of the Affymetrix GCOS software), with green lines indicating twofold, threefold, 10-fold, and 30-fold differential expression between the two samples. The arrow points to signals from three independent IGF2 <t>mRNAs</t> probes. ( B ) Genes that are changed (either up-regulated or down-regulated) more than twofold in MHH-ES-1 cells transfected with a miR-483-5p mimic compared with scrambled control. ( C ) Time course of IGF2 mRNA expression after transfection of MHH-ES-1 with a miR-483-5p mimic, as measured by qRT–PCR. Expression is relative to levels at day 0. ( D ) Induction of IGF2 mRNA by miR-483-5p in two human sarcoma cell lines (MMH-ES-1 and RD) and two mouse sarcoma lines (F4328 and F4864), as measured by qRT–PCR. The asterisk denotes a significant difference ( P
    Mrnas, supplied by Unigene, used in various techniques. Bioz Stars score: 93/100, based on 265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore mrna
    Splicing analysis by <t>RT–PCR</t> of specific genes in U5 snRNA loop 1 mutants. Inverted images of ethidium bromide stained agarose gels. Poly(A) + <t>mRNA</t> isolated from total RNA was subjected to RT–PCR. The genes analyzed are listed to the left of each panel. The unspliced pre-mRNA (P) and spliced mRNA (M) are indicated on the right of each panel. Control reactions that did not include reverse transcription of poly(A) + RNA are indicated above each lane with a minus (–).
    Mrna, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 2985 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TriLink mrna delivery cas9 mrna
    Splicing analysis by <t>RT–PCR</t> of specific genes in U5 snRNA loop 1 mutants. Inverted images of ethidium bromide stained agarose gels. Poly(A) + <t>mRNA</t> isolated from total RNA was subjected to RT–PCR. The genes analyzed are listed to the left of each panel. The unspliced pre-mRNA (P) and spliced mRNA (M) are indicated on the right of each panel. Control reactions that did not include reverse transcription of poly(A) + RNA are indicated above each lane with a minus (–).
    Mrna Delivery Cas9 Mrna, supplied by TriLink, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mrna  (Qiagen)
    99
    Qiagen mrna
    Validation of the expression patterns of selected oat genes by qRT-PCR. ( A ) Changes in the relative <t>mRNA</t> levels of 18 selected genes as determined by <t>RNA-Seq.</t> The Y axis indicates the relative fold change in transcript abundance under conditions of salt stress in relation to the control. S: ‘Huazao-2’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. T: ‘Hanyou-5’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. ( B ) Changes in the relative mRNA levels of 18 selected genes as determined by qRT-PCR. The qRT-PCR experiment was carried out with three biological replicates, and the actin gene was used as an internal control. The results are presented as target/reference ratios normalized by the internal control.
    Mrna, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 27082 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    RGEN-induced Prkdc gene disruption in mice. ( A ) Representative T7E1 assays demonstrating mutagenesis efficiencies of Cas9 mRNA plus Prkdc -specific sgRNA that were delivered via intracytoplasmic injection into one-cell-stage mouse embryos. Numbers indicate

    Journal: Genome Research

    Article Title: Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases

    doi: 10.1101/gr.163394.113

    Figure Lengend Snippet: RGEN-induced Prkdc gene disruption in mice. ( A ) Representative T7E1 assays demonstrating mutagenesis efficiencies of Cas9 mRNA plus Prkdc -specific sgRNA that were delivered via intracytoplasmic injection into one-cell-stage mouse embryos. Numbers indicate

    Article Snippet: Cas9 mRNA and sgRNAs in M2 medium (Sigma-Aldrich) were injected into the cytoplasm of fertilized eggs with well-recognized pronuclei using a Piezo-driven micromanipulator (Prime Tech).

    Techniques: Mouse Assay, Mutagenesis, Injection

    Effects of chitosan nanofibers on the induction of osteoblast differentiation-related OPN, OCN, and ALP gene expression. Notes: Human osteoblast-like MG63 cells were seeded on chitosan nanofiber scaffolds for 1, 3, and 5 days. ( A ) Analyses of OPN, OCN, and ALP mRNAs were conducted using RT-PCR; β-actin mRNA was analyzed as an internal control. The DNA bands were quantified and statistically analyzed ( B – D ). Each value represents the mean ± standard error of the mean from four independent experiments. * indicates that values significantly differed from the respective control, P

    Journal: International Journal of Nanomedicine

    Article Title: Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

    doi: 10.2147/IJN.S68012

    Figure Lengend Snippet: Effects of chitosan nanofibers on the induction of osteoblast differentiation-related OPN, OCN, and ALP gene expression. Notes: Human osteoblast-like MG63 cells were seeded on chitosan nanofiber scaffolds for 1, 3, and 5 days. ( A ) Analyses of OPN, OCN, and ALP mRNAs were conducted using RT-PCR; β-actin mRNA was analyzed as an internal control. The DNA bands were quantified and statistically analyzed ( B – D ). Each value represents the mean ± standard error of the mean from four independent experiments. * indicates that values significantly differed from the respective control, P

    Article Snippet: Oligonucleotides for the PCR analyses of these mRNAs were designed and synthesized by Takara Bio Inc. (Otsu, Japan).

    Techniques: ALP Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    Host cell mRNA synthesis and translation are unaffected by compound JMN3-003. A ) Relative TaqMan RT-PCR-based quantitation of three unstable cellular mRNAs (MCL1, ASB7, MKP-1) after exposure of cells to JMN3-003 for six hours. Controls were treated with Actinomycine D (Act D) for comparison. C T values are expressed relative to vehicle-treated samples and reflect averages of three independent experiments, each analyzed in triplicate, ± SD. B–D ) Expression of virus-encoded but not host cell or plasmid-encoded viral proteins is blocked by JMN3-003. Immunodetection of transiently expressed MeV-F ( B ), virus-encoded MeV-F ( C ), and virus-encoded influenza A/WSN M2 ( D ) in cell lysates after incubation of cells in the presence of compound or vehicle only (DMSO) for 30 hours. As internal cellular standard, membranes were probed for GAPDH in parallel. Numbers correspond to average densitometric quantitations ± SD of three experiments, representative immunoblots are shown. (ND: not determined).

    Journal: PLoS ONE

    Article Title: Potent Host-Directed Small-Molecule Inhibitors of Myxovirus RNA-Dependent RNA-Polymerases

    doi: 10.1371/journal.pone.0020069

    Figure Lengend Snippet: Host cell mRNA synthesis and translation are unaffected by compound JMN3-003. A ) Relative TaqMan RT-PCR-based quantitation of three unstable cellular mRNAs (MCL1, ASB7, MKP-1) after exposure of cells to JMN3-003 for six hours. Controls were treated with Actinomycine D (Act D) for comparison. C T values are expressed relative to vehicle-treated samples and reflect averages of three independent experiments, each analyzed in triplicate, ± SD. B–D ) Expression of virus-encoded but not host cell or plasmid-encoded viral proteins is blocked by JMN3-003. Immunodetection of transiently expressed MeV-F ( B ), virus-encoded MeV-F ( C ), and virus-encoded influenza A/WSN M2 ( D ) in cell lysates after incubation of cells in the presence of compound or vehicle only (DMSO) for 30 hours. As internal cellular standard, membranes were probed for GAPDH in parallel. Numbers correspond to average densitometric quantitations ± SD of three experiments, representative immunoblots are shown. (ND: not determined).

    Article Snippet: Quantitative TaqMan RT-PCR was again achieved using the TaqMan Fast Master Mix (Applied Biosystems) combined with proprietary primer and probe sets specific for Induced myeloid leukemia cell differentiation protein 1- (MCL1), MAPK phosphatase 1 (MKP1), and ankyrin repeat and SOCS box-containing protein 7- (ASB7) encoding mRNAs (Applied Biosystems).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitation Assay, Activated Clotting Time Assay, Expressing, Plasmid Preparation, Immunodetection, Incubation, Western Blot

    Expression profiles and enrichment Analysis of mRNAs/lncRNAs regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p

    Journal: Scientific Reports

    Article Title: Let-7e modulates the inflammatory response in vascular endothelial cells through ceRNA crosstalk

    doi: 10.1038/srep42498

    Figure Lengend Snippet: Expression profiles and enrichment Analysis of mRNAs/lncRNAs regulated by let-7e. (a) The percentage of down-regulated mRNAs (lncRNAs) in all regulated mRNAs (lncRNAs) in HUVECs treated by let-7e mimic or inhibitor. ★ p

    Article Snippet: Expression data of mRNAs and lncRNAs were filtered and normalized (ArrayExpress accession number: E-MTAB-4971), and then the lncRNAs and mRNAs regulated by let-7e were identified using GeneSpring GX 11.5 (Agilent, CA).

    Techniques: Expressing

    Maternal Vangl2 and aPKC depletion disrupts the pattern of expression of specific zygotic genes. ( A ) Relative expression levels of Siamois , Xnr3 and Foxi1e mRNAs in control (Uninj) and Vangl2-depleted (VG–) embryos at the late blastula (st9) and

    Journal: Development (Cambridge, England)

    Article Title: The roles of maternal Vangl2 and aPKC in Xenopus oocyte and embryo patterning

    doi: 10.1242/dev.068866

    Figure Lengend Snippet: Maternal Vangl2 and aPKC depletion disrupts the pattern of expression of specific zygotic genes. ( A ) Relative expression levels of Siamois , Xnr3 and Foxi1e mRNAs in control (Uninj) and Vangl2-depleted (VG–) embryos at the late blastula (st9) and

    Article Snippet: The roles of PCP and ABP pathway components have not been examined in vertebrate oocytes, although real-time RT-PCR analysis shows that their mRNAs (including Vangl2, Celsr1, Disheveled, frizzled, prickle, PTK7, aPKC, scribble, Par3 and Par6 ) are expressed (data not shown).

    Techniques: Expressing

    Expression of HSI2 , HSL1 , and seed maturation genes in Col-0 and KK seedlings. Total RNAs were isolated from 4-, 5-, 7-, and 9-day-old Col-0 and KK seedlings grown on medium containing 1% sucrose. The levels of various mRNAs were determined by quantitative real-time RT-PCR using ACTIN2 mRNA as an internal reference. Results represent the average from two independent isolations of RNA ± SD.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Two B3 domain transcriptional repressors prevent sugar-inducible expression of seed maturation genes in Arabidopsis seedlings

    doi: 10.1073/pnas.0607940104

    Figure Lengend Snippet: Expression of HSI2 , HSL1 , and seed maturation genes in Col-0 and KK seedlings. Total RNAs were isolated from 4-, 5-, 7-, and 9-day-old Col-0 and KK seedlings grown on medium containing 1% sucrose. The levels of various mRNAs were determined by quantitative real-time RT-PCR using ACTIN2 mRNA as an internal reference. Results represent the average from two independent isolations of RNA ± SD.

    Article Snippet: Total RNA was isolated from plants by using an RNeasy plant mini kit (Qiagen, Valencia, CA), and levels of various mRNAs were determined by quantitative real-time RT-PCR using an iCycler iQ with iQ SYBR green supermix (Bio-Rad, Hercules, CA) as described previously ( ).

    Techniques: Expressing, Isolation, Quantitative RT-PCR

    Effects of the concentration of sucrose in the medium on the expression of various genes in seedlings. Total RNA was isolated from 5-day-old Col-0 and KK seedlings grown on medium containing 0, 10, 30, and 90 mM sucrose. The levels of various mRNAs were determined by quantitative real-time RT-PCR using ACTIN2 mRNA as an internal reference. Results represent the average of data from two independent isolations of RNA ± SD.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Two B3 domain transcriptional repressors prevent sugar-inducible expression of seed maturation genes in Arabidopsis seedlings

    doi: 10.1073/pnas.0607940104

    Figure Lengend Snippet: Effects of the concentration of sucrose in the medium on the expression of various genes in seedlings. Total RNA was isolated from 5-day-old Col-0 and KK seedlings grown on medium containing 0, 10, 30, and 90 mM sucrose. The levels of various mRNAs were determined by quantitative real-time RT-PCR using ACTIN2 mRNA as an internal reference. Results represent the average of data from two independent isolations of RNA ± SD.

    Article Snippet: Total RNA was isolated from plants by using an RNeasy plant mini kit (Qiagen, Valencia, CA), and levels of various mRNAs were determined by quantitative real-time RT-PCR using an iCycler iQ with iQ SYBR green supermix (Bio-Rad, Hercules, CA) as described previously ( ).

    Techniques: Concentration Assay, Expressing, Isolation, Quantitative RT-PCR

    Schematic representation of iPSC derivation protocol and the generation of iPSCs from HSF and HUVECs. (A) On day 0, the parent cells, HSF and HVECs cells were treated separately with Stemcircle, a plasmid containing Oct4, Nanog, Sox2 and Lin28 for 24 hours and subsequently, a cocktail of mRNAs containing Oct4, Sox2, Klf4, cMyc, Lin28 for up to day 12. The colonies were allowed to expand in between day 13 and 16. On day 17, the fully expanded colonies were picked and grown. HSF-Human skin fibroblast, HUVECs-Human umbilical vein endothelial cells, D-day. (B1) Phage contrast microscopic image of iPSC-derived from HSF (hf-iPSCs). (B2) Live staining of Tra1-60+ cells from hf-iPSCs. (C1) Phage contrast microscopic image of iPSC-derived from HUVECs (he-iPSCs). (C2) Live staining of Tra1-60+ cells from he-iPSCs. (D) Transfection efficiency of iPSCs generated from HSF by flow cytometry analysis at passage 0 (P0). The data show 4% of cells were positive for either Oct4 or Sox2. (E) Quantification of Tra1-60+ cells by flow cytometry at P9. The data show 73.2% of cells are Tra1-60+. Data shown are representative of three independent experiments, **p

    Journal: PLoS ONE

    Article Title: Generation of Functional Cardiomyocytes from Efficiently Generated Human iPSCs and a Novel Method of Measuring Contractility

    doi: 10.1371/journal.pone.0134093

    Figure Lengend Snippet: Schematic representation of iPSC derivation protocol and the generation of iPSCs from HSF and HUVECs. (A) On day 0, the parent cells, HSF and HVECs cells were treated separately with Stemcircle, a plasmid containing Oct4, Nanog, Sox2 and Lin28 for 24 hours and subsequently, a cocktail of mRNAs containing Oct4, Sox2, Klf4, cMyc, Lin28 for up to day 12. The colonies were allowed to expand in between day 13 and 16. On day 17, the fully expanded colonies were picked and grown. HSF-Human skin fibroblast, HUVECs-Human umbilical vein endothelial cells, D-day. (B1) Phage contrast microscopic image of iPSC-derived from HSF (hf-iPSCs). (B2) Live staining of Tra1-60+ cells from hf-iPSCs. (C1) Phage contrast microscopic image of iPSC-derived from HUVECs (he-iPSCs). (C2) Live staining of Tra1-60+ cells from he-iPSCs. (D) Transfection efficiency of iPSCs generated from HSF by flow cytometry analysis at passage 0 (P0). The data show 4% of cells were positive for either Oct4 or Sox2. (E) Quantification of Tra1-60+ cells by flow cytometry at P9. The data show 73.2% of cells are Tra1-60+. Data shown are representative of three independent experiments, **p

    Article Snippet: Then the cells were followed by a cocktail of mRNAs (Stemgent) containing Oct4, Sox2, Klf4, cMyc and Lin28 every day for 11 days using Lipofectamine (Invitrogen) as a transfection agent.

    Techniques: Plasmid Preparation, Derivative Assay, Staining, Transfection, Generated, Flow Cytometry, Cytometry

    miR-483-5p induces promoter-specific IGF2 transcription. ( A ) Scatter plot (log) of mRNA profiles of MHH-ES-1 cells transfected with miR-483-5p ( Y -axis) versus scrambled control ( X -axis). Transcripts expressed in both samples (red), one sample (blue), or neither sample (yellow) are shown (as determined by default settings of the Affymetrix GCOS software), with green lines indicating twofold, threefold, 10-fold, and 30-fold differential expression between the two samples. The arrow points to signals from three independent IGF2 mRNAs probes. ( B ) Genes that are changed (either up-regulated or down-regulated) more than twofold in MHH-ES-1 cells transfected with a miR-483-5p mimic compared with scrambled control. ( C ) Time course of IGF2 mRNA expression after transfection of MHH-ES-1 with a miR-483-5p mimic, as measured by qRT–PCR. Expression is relative to levels at day 0. ( D ) Induction of IGF2 mRNA by miR-483-5p in two human sarcoma cell lines (MMH-ES-1 and RD) and two mouse sarcoma lines (F4328 and F4864), as measured by qRT–PCR. The asterisk denotes a significant difference ( P

    Journal: Genes & Development

    Article Title: The IGF2 intronic miR-483 selectively enhances transcription from IGF2 fetal promoters and enhances tumorigenesis

    doi: 10.1101/gad.224170.113

    Figure Lengend Snippet: miR-483-5p induces promoter-specific IGF2 transcription. ( A ) Scatter plot (log) of mRNA profiles of MHH-ES-1 cells transfected with miR-483-5p ( Y -axis) versus scrambled control ( X -axis). Transcripts expressed in both samples (red), one sample (blue), or neither sample (yellow) are shown (as determined by default settings of the Affymetrix GCOS software), with green lines indicating twofold, threefold, 10-fold, and 30-fold differential expression between the two samples. The arrow points to signals from three independent IGF2 mRNAs probes. ( B ) Genes that are changed (either up-regulated or down-regulated) more than twofold in MHH-ES-1 cells transfected with a miR-483-5p mimic compared with scrambled control. ( C ) Time course of IGF2 mRNA expression after transfection of MHH-ES-1 with a miR-483-5p mimic, as measured by qRT–PCR. Expression is relative to levels at day 0. ( D ) Induction of IGF2 mRNA by miR-483-5p in two human sarcoma cell lines (MMH-ES-1 and RD) and two mouse sarcoma lines (F4328 and F4864), as measured by qRT–PCR. The asterisk denotes a significant difference ( P

    Article Snippet: Human primary tissues used to assess the expression of IGF2 promoter–specific mRNAs and miR-483-5p were obtained from Stratagene and Zyagen, as described in the Supplemental Material.

    Techniques: Transfection, Software, Expressing, Quantitative RT-PCR

    miR-483-5p localizes to the nucleus and binds the 5′ UTR of nuclear IGF2 P3 mRNA to induce gene activation. ( A ) FISH showing nuclear localization of endogenous miR-483-5p (FITC; green) using a specific LNA probe against the miRNA. The nucleus was stained with DAPI (blue). The small nuclear RNA U6 is shown as positive control for nuclear signal. LNA probes detecting the miRNA-483-5p mutant and the WPMY-1 fibroblast cell line that lacks IGF2 expression were used as negative controls. Two unrelated miRNAs (miR-675 and miR-31) lack apparent nuclear localization. ( B ) Schematic representation of potential miR-483-binding sites within the 5′ UTR region of IGF2 P3 transcript, including miR-483-5p (site 1 and site 2) and miR-483-3p. Fragments subjected to pull-down assays are shown with letters (a–e), with amplification primers denoted by arrowheads. ( C ) Biotinylated miRNA affinity pull-down was conducted using streptavidin beads with nuclear and cytosolic extracts of cells transfected with different biotinylated miRNA baits followed by qRT–PCR analysis for the presence of specific regions of IGF2 mRNA. The miRNA baits are as follows: biotinylated miR-483-5p (1), biotinylated miR-483-5p mutant (2), sense strand biotinylated miR-483-5p (3), biotinylated miR-483-3p (4), and nonbiotinylated miR-483-5p (5). PCR-amplified fragments are as follows: P3 promoter (region a), P3 5′ UTR (region b), and the intronic region between the P3 5′ UTR and the P4 5′ UTR (region c) that represents the spliced intron sequence. Region d identifies the spliced mature P3 mRNA, region e identifies total IGF2 mRNAs, and region f identifies GAPDH mRNA, which serves as a control. Cyto and Nuc represent pull-down from cytoplasmic and nuclear fractions, respectively. ( D ) Suppression of IGF2 mRNA induction in MHH-ES-1 cells by miR-483-5p (vs. scrambled control) following transfection of a 2′- O -methyl antisense protector RNA that specifically targets site 2. The 2′- O -methyl antisense RNAs target site 1 alone (α Site1), site 2 alone (α Site2), site 1 plus site 2 (α Site1 + α Site2), GAPDH 5′ UTR (α GAPDH ), or mutant site 2 (α Site2 mutant). The asterisk denotes a significant difference ( P

    Journal: Genes & Development

    Article Title: The IGF2 intronic miR-483 selectively enhances transcription from IGF2 fetal promoters and enhances tumorigenesis

    doi: 10.1101/gad.224170.113

    Figure Lengend Snippet: miR-483-5p localizes to the nucleus and binds the 5′ UTR of nuclear IGF2 P3 mRNA to induce gene activation. ( A ) FISH showing nuclear localization of endogenous miR-483-5p (FITC; green) using a specific LNA probe against the miRNA. The nucleus was stained with DAPI (blue). The small nuclear RNA U6 is shown as positive control for nuclear signal. LNA probes detecting the miRNA-483-5p mutant and the WPMY-1 fibroblast cell line that lacks IGF2 expression were used as negative controls. Two unrelated miRNAs (miR-675 and miR-31) lack apparent nuclear localization. ( B ) Schematic representation of potential miR-483-binding sites within the 5′ UTR region of IGF2 P3 transcript, including miR-483-5p (site 1 and site 2) and miR-483-3p. Fragments subjected to pull-down assays are shown with letters (a–e), with amplification primers denoted by arrowheads. ( C ) Biotinylated miRNA affinity pull-down was conducted using streptavidin beads with nuclear and cytosolic extracts of cells transfected with different biotinylated miRNA baits followed by qRT–PCR analysis for the presence of specific regions of IGF2 mRNA. The miRNA baits are as follows: biotinylated miR-483-5p (1), biotinylated miR-483-5p mutant (2), sense strand biotinylated miR-483-5p (3), biotinylated miR-483-3p (4), and nonbiotinylated miR-483-5p (5). PCR-amplified fragments are as follows: P3 promoter (region a), P3 5′ UTR (region b), and the intronic region between the P3 5′ UTR and the P4 5′ UTR (region c) that represents the spliced intron sequence. Region d identifies the spliced mature P3 mRNA, region e identifies total IGF2 mRNAs, and region f identifies GAPDH mRNA, which serves as a control. Cyto and Nuc represent pull-down from cytoplasmic and nuclear fractions, respectively. ( D ) Suppression of IGF2 mRNA induction in MHH-ES-1 cells by miR-483-5p (vs. scrambled control) following transfection of a 2′- O -methyl antisense protector RNA that specifically targets site 2. The 2′- O -methyl antisense RNAs target site 1 alone (α Site1), site 2 alone (α Site2), site 1 plus site 2 (α Site1 + α Site2), GAPDH 5′ UTR (α GAPDH ), or mutant site 2 (α Site2 mutant). The asterisk denotes a significant difference ( P

    Article Snippet: Human primary tissues used to assess the expression of IGF2 promoter–specific mRNAs and miR-483-5p were obtained from Stratagene and Zyagen, as described in the Supplemental Material.

    Techniques: Activation Assay, Fluorescence In Situ Hybridization, Staining, Positive Control, Mutagenesis, Expressing, Binding Assay, Amplification, Transfection, Quantitative RT-PCR, Polymerase Chain Reaction, Sequencing

    miR-483-5p enhances the interaction of IGF2 P3 mRNA with the RNA helicase A (DHX9). ( A ) Silver-stained gel of electrophoresed proteins pulled down using the biotinylated 5′ UTR of IGF2 P3 mRNA following transfection of cells with scrambled miRNA control (lanes 1 , 2 ), miR-483-5p (lane 3 ), miR-483-5p + antisense protector targeting site 2 (lane 4 ), and miR-483-5p + antisense mutated protector (lane 5 ). The pull-down experiments were performed using biotin-labeled IGF2 P3 mRNA 5′ UTR (L), with the identical, unlabeled IGF2 mRNA fragment serving as negative control (U). The protein that is differentially bound to IGF2 P3 mRNA 5′ UTR in the presence of miR-483-5p and in the absence of the inhibitory protector (red box in lane 3 , L) was sequenced and found to be DHX9. ( B ) Western blot confirmation of DHX9 pull-down by IGF2 P3 5′ UTR RNA. DHX9 content for each input sample is shown. Lamin A/C serves as the loading control. ( C ) Number of DHX9 peptides associated with miR-483-5p or control miRNA baits, measured by mass spectrometry. Black and white bars represent unique peptide numbers and total peptide numbers identified, respectively. ( D ) Immunoprecipitation of endogenous DHX9 in MHH-ES-1 cells and those transfected with antisense protector targeting site 2 (α Site2) or antisense mutated protector (α Site2 mutant). Four mRNAs— IGF2 P2, P3, P4 and H19—were assayed by qRT–PCR. ( E ) siRNA-mediated knockdown of DHX9 abrogates the induction of IGF2 by miR-483-5p and reduces endogenous IGF2 mRNA in the absence of ectopic miR-483-5p. The asterisk denotes a significant difference ( P

    Journal: Genes & Development

    Article Title: The IGF2 intronic miR-483 selectively enhances transcription from IGF2 fetal promoters and enhances tumorigenesis

    doi: 10.1101/gad.224170.113

    Figure Lengend Snippet: miR-483-5p enhances the interaction of IGF2 P3 mRNA with the RNA helicase A (DHX9). ( A ) Silver-stained gel of electrophoresed proteins pulled down using the biotinylated 5′ UTR of IGF2 P3 mRNA following transfection of cells with scrambled miRNA control (lanes 1 , 2 ), miR-483-5p (lane 3 ), miR-483-5p + antisense protector targeting site 2 (lane 4 ), and miR-483-5p + antisense mutated protector (lane 5 ). The pull-down experiments were performed using biotin-labeled IGF2 P3 mRNA 5′ UTR (L), with the identical, unlabeled IGF2 mRNA fragment serving as negative control (U). The protein that is differentially bound to IGF2 P3 mRNA 5′ UTR in the presence of miR-483-5p and in the absence of the inhibitory protector (red box in lane 3 , L) was sequenced and found to be DHX9. ( B ) Western blot confirmation of DHX9 pull-down by IGF2 P3 5′ UTR RNA. DHX9 content for each input sample is shown. Lamin A/C serves as the loading control. ( C ) Number of DHX9 peptides associated with miR-483-5p or control miRNA baits, measured by mass spectrometry. Black and white bars represent unique peptide numbers and total peptide numbers identified, respectively. ( D ) Immunoprecipitation of endogenous DHX9 in MHH-ES-1 cells and those transfected with antisense protector targeting site 2 (α Site2) or antisense mutated protector (α Site2 mutant). Four mRNAs— IGF2 P2, P3, P4 and H19—were assayed by qRT–PCR. ( E ) siRNA-mediated knockdown of DHX9 abrogates the induction of IGF2 by miR-483-5p and reduces endogenous IGF2 mRNA in the absence of ectopic miR-483-5p. The asterisk denotes a significant difference ( P

    Article Snippet: Human primary tissues used to assess the expression of IGF2 promoter–specific mRNAs and miR-483-5p were obtained from Stratagene and Zyagen, as described in the Supplemental Material.

    Techniques: Staining, Transfection, Labeling, Negative Control, Western Blot, Mass Spectrometry, Immunoprecipitation, Mutagenesis, Quantitative RT-PCR

    Splicing analysis by RT–PCR of specific genes in U5 snRNA loop 1 mutants. Inverted images of ethidium bromide stained agarose gels. Poly(A) + mRNA isolated from total RNA was subjected to RT–PCR. The genes analyzed are listed to the left of each panel. The unspliced pre-mRNA (P) and spliced mRNA (M) are indicated on the right of each panel. Control reactions that did not include reverse transcription of poly(A) + RNA are indicated above each lane with a minus (–).

    Journal: Nucleic Acids Research

    Article Title: Mutations in U5 snRNA loop 1 influence the splicing of different genes in vivo

    doi:

    Figure Lengend Snippet: Splicing analysis by RT–PCR of specific genes in U5 snRNA loop 1 mutants. Inverted images of ethidium bromide stained agarose gels. Poly(A) + mRNA isolated from total RNA was subjected to RT–PCR. The genes analyzed are listed to the left of each panel. The unspliced pre-mRNA (P) and spliced mRNA (M) are indicated on the right of each panel. Control reactions that did not include reverse transcription of poly(A) + RNA are indicated above each lane with a minus (–).

    Article Snippet: For RT–PCR, poly(A)+ mRNA was isolated from 200 µg of total RNA prepared as above from cells grown at 37°C for 12 h using the GenElute mRNA miniprep kit (Sigma).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Staining, Isolation

    Validation of the expression patterns of selected oat genes by qRT-PCR. ( A ) Changes in the relative mRNA levels of 18 selected genes as determined by RNA-Seq. The Y axis indicates the relative fold change in transcript abundance under conditions of salt stress in relation to the control. S: ‘Huazao-2’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. T: ‘Hanyou-5’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. ( B ) Changes in the relative mRNA levels of 18 selected genes as determined by qRT-PCR. The qRT-PCR experiment was carried out with three biological replicates, and the actin gene was used as an internal control. The results are presented as target/reference ratios normalized by the internal control.

    Journal: Scientific Reports

    Article Title: Comparative Transcriptional Profiling and Physiological Responses of Two Contrasting Oat Genotypes under Salt Stress

    doi: 10.1038/s41598-018-34505-5

    Figure Lengend Snippet: Validation of the expression patterns of selected oat genes by qRT-PCR. ( A ) Changes in the relative mRNA levels of 18 selected genes as determined by RNA-Seq. The Y axis indicates the relative fold change in transcript abundance under conditions of salt stress in relation to the control. S: ‘Huazao-2’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. T: ‘Hanyou-5’ under normal growth conditions (1) and after 2 (2), 4 (3), 8 (4), 12 (5), and 24 (6) h of salt stress treatment. ( B ) Changes in the relative mRNA levels of 18 selected genes as determined by qRT-PCR. The qRT-PCR experiment was carried out with three biological replicates, and the actin gene was used as an internal control. The results are presented as target/reference ratios normalized by the internal control.

    Article Snippet: RNA extraction and double-stranded cDNA synthesis Total RNA was extracted from whole plants using the RNeasy Plant Mini Kit (Qiagen) according to the manufacturer’s instructions. mRNA was isolated from the qualified total RNA using the Oligotex mRNA Mini Kit (Qiagen). mRNA extracted from the salt-stressed oat seedlings after different durations was prepared for cDNA synthesis.

    Techniques: Expressing, Quantitative RT-PCR, RNA Sequencing Assay