qrt-pcr assays Search Results


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  • 99
    Thermo Fisher mirvana qrt pcr mirna detection kit
    miR-155 promotes production of TNF-α and IL-1β in human PBMCs. ( A ) Fold changes of miR-155 expression were determined by <t>qRT-PCR</t> after the transfection of miR-control, or miR-155 mimics at 25 or 50 nM for 24 h; ( B ) and ( C ) Expression of TNF-α and IL-1β in supernatant of PBMCs 24 h post LPS treatment, or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM <t>miRNA</t> control. Results were expressed as pg/mL by an ELISA test; and ( D ) mRNA expression of TNF-α (group 1) and IL-1β (group 2) in PBMCs 24 h after LPS treatment or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM miRNA control. All results are the average of three independent experiments. Statistical significance is shown.
    Mirvana Qrt Pcr Mirna Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 981 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad qrt pcr
    After osteogenic induction, total RNA was extracted from all groups and expression of RUNX 2 and OCN was assessed by <t>qRT-PCR.</t> The expression of RUNX 2 and OCN of hUSCs (P4) from the children group was higher than that from the elder group (* P
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    Thermo Fisher qrt pcr
    After osteogenic induction, total RNA was extracted from all groups and expression of RUNX 2 and OCN was assessed by <t>qRT-PCR.</t> The expression of RUNX 2 and OCN of hUSCs (P4) from the children group was higher than that from the elder group (* P
    Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene qrt pcr
    After osteogenic induction, total RNA was extracted from all groups and expression of RUNX 2 and OCN was assessed by <t>qRT-PCR.</t> The expression of RUNX 2 and OCN of hUSCs (P4) from the children group was higher than that from the elder group (* P
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    Thermo Fisher superscript iii platinum one step qrt pcr kit
    Effect of NLRC5 KD on differentially expressed immune genes during H5N2 infection. NLRC5 siRNA and scramble control transfected MQ-NCSU cells were infected with H5N2 virus. Total RNA was collected at 24 hpi and assayed by the RT 2 Profiler <t>PCR</t> Array system. Selected genes taking part in innate and adaptive immune response are shown as log 2 transform of fold-change expression relative to the gene expression in the mock-transfected cells. Data represent an average of <t>three</t> biological replicates.
    Superscript Iii Platinum One Step Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1710 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Toyobo qrt pcr
    Effect of NLRC5 KD on differentially expressed immune genes during H5N2 infection. NLRC5 siRNA and scramble control transfected MQ-NCSU cells were infected with H5N2 virus. Total RNA was collected at 24 hpi and assayed by the RT 2 Profiler <t>PCR</t> Array system. Selected genes taking part in innate and adaptive immune response are shown as log 2 transform of fold-change expression relative to the gene expression in the mock-transfected cells. Data represent an average of <t>three</t> biological replicates.
    Qrt Pcr, supplied by Toyobo, used in various techniques. Bioz Stars score: 94/100, based on 494 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii platinum sybr green one step qrt pcr kit
    In vitro and ex vivo transcriptional regulation of chIFIT5 gene. ( A ) ChIFN-β itself, or produced by different stimuli, can initiate JAK-STAT signalling pathway and culminate in the induction of hundreds of ISGs. IFIT family of genes is one such antiviral effector of innate immune responses. ( B ) Quantitation of chIFIT5 mRNA in cells stimulated with 1000 U of chIFN-β, 10 μg/mL of LPS, 5 μg/mL of poly I:C or 1 MOI of NDV for 24 hours before RNA extraction and analysis for <t>qRT-PCR</t> using primers specific for the chIFIT5 gene. ( C–F ) Total cellular RNA was extracted from CEF cells at 1, 2, 4, 8, 16, and 24 hours post-NDV infection and was subjected to quantitation of mRNA for chIFIT5 ( C ), viral mRNA ( D ), chMx ( E ) and chIFN-β ( F , G ) RNA collected from seven organs infected or not with H9N2 influenza viruses were used to determine the level of chIFIT5 mRNA and M gene of the virus. Fold change induction in all experiments was determined by 2 −∆∆CT algorithm and data presented is average of <t>three</t> independent experiments.
    Superscript Iii Platinum Sybr Green One Step Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1742 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genecopoeia all in one mirna qrt pcr detection kit
    TAp63 modulates the expression of miR-133b. HT-29 and SW-620 cells were transfected with either an empty vector or with TAp63 for 72 h. ( A ) Endogenous TAp63 levels were assessed by <t>RT–PCR.</t> ( B ) The expression level of TAp63 is shown in the western blotting. ( C ). Endogenous TAp63 levels were again confirmed by <t>qRT–PCR.</t> As the figure shows, HT-29 and SW-620 cells transfected with TAp63 expressing vector induced a significant increase in TAp63 expression. ( D ) Endogenous miR-133b levels were assessed by qRT–PCR. After transfection with a TAp63-expressing vector, HT-29 and SW-620 cells showed a significant increase in miR-133b expression. ( E ) Chromatin immunoprecipitation (ChIP) experiment showed that p63 is able to bind the p53 RE-III site but not p53 RE-I and p53 RE-II. ( F ) Insertion of miR-133b promoter region in a luciferase reporter gene leads to increased luciferase activity in the presence of TAp63 in HT-29 and SW-620 cells. Mutation of the RE-III p53-binding site abolished TAp63-mediated luciferase activity. ( G ) The miR-133b promoter region containing the three p53 consensus sites (p53RE). All qRT–PCR results were relative to U6-snRNA or GAPDH and were normalised to the expression of miR-133b or TAp63 in HT-29 cells. Data represent the means±s.d. of three different experiments analysed in triplicate. * P
    All In One Mirna Qrt Pcr Detection Kit, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 94/100, based on 1519 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Eppendorf AG qrt pcr
    TAp63 modulates the expression of miR-133b. HT-29 and SW-620 cells were transfected with either an empty vector or with TAp63 for 72 h. ( A ) Endogenous TAp63 levels were assessed by <t>RT–PCR.</t> ( B ) The expression level of TAp63 is shown in the western blotting. ( C ). Endogenous TAp63 levels were again confirmed by <t>qRT–PCR.</t> As the figure shows, HT-29 and SW-620 cells transfected with TAp63 expressing vector induced a significant increase in TAp63 expression. ( D ) Endogenous miR-133b levels were assessed by qRT–PCR. After transfection with a TAp63-expressing vector, HT-29 and SW-620 cells showed a significant increase in miR-133b expression. ( E ) Chromatin immunoprecipitation (ChIP) experiment showed that p63 is able to bind the p53 RE-III site but not p53 RE-I and p53 RE-II. ( F ) Insertion of miR-133b promoter region in a luciferase reporter gene leads to increased luciferase activity in the presence of TAp63 in HT-29 and SW-620 cells. Mutation of the RE-III p53-binding site abolished TAp63-mediated luciferase activity. ( G ) The miR-133b promoter region containing the three p53 consensus sites (p53RE). All qRT–PCR results were relative to U6-snRNA or GAPDH and were normalised to the expression of miR-133b or TAp63 in HT-29 cells. Data represent the means±s.d. of three different experiments analysed in triplicate. * P
    Qrt Pcr, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 94/100, based on 445 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii first strand synthesis supermix for qrt pcr
    Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by <t>qRT-PCR.</t> (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of <t>three</t> biological replicates. ** P
    Superscript Iii First Strand Synthesis Supermix For Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies qrt pcr
    Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by <t>qRT-PCR.</t> (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of <t>three</t> biological replicates. ** P
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    tiangen biotech co qrt pcr
    Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by <t>qRT-PCR.</t> (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of <t>three</t> biological replicates. ** P
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    Illumina Inc qrt pcr
    Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by <t>qRT-PCR.</t> (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of <t>three</t> biological replicates. ** P
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    Thermo Fisher single cell to ct qrt pcr kit
    miR-370 regulates MGMT expression in raji cells. a <t>qRT-PCR</t> demonstrated that miR-370 inhibitors increased expression of MGMT at mRNA level, while miR-370 mimics suppressed MGMT mRNA expression. b The result of MGMT protein level examined by western blot was similar with ( a ) after being transfected with miR-370 inhibitors or mimics
    Single Cell To Ct Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vazyme Biotech Co qrt pcr
    miR-370 regulates MGMT expression in raji cells. a <t>qRT-PCR</t> demonstrated that miR-370 inhibitors increased expression of MGMT at mRNA level, while miR-370 mimics suppressed MGMT mRNA expression. b The result of MGMT protein level examined by western blot was similar with ( a ) after being transfected with miR-370 inhibitors or mimics
    Qrt Pcr, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 92/100, based on 301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Meridian Life Science qrt pcr
    miR-370 regulates MGMT expression in raji cells. a <t>qRT-PCR</t> demonstrated that miR-370 inhibitors increased expression of MGMT at mRNA level, while miR-370 mimics suppressed MGMT mRNA expression. b The result of MGMT protein level examined by western blot was similar with ( a ) after being transfected with miR-370 inhibitors or mimics
    Qrt Pcr, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 92/100, based on 209 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii platinum two step qrt pcr kit
    xnp1 gene expression is induced by mitomycin C treatment. Results shown are for <t>qRT-PCR</t> analysis of xnp1 gene expression before treatment with mitomycin C (0 h) and 1 h and 2.5 h after induction. For each gene, time zero was arbitrarily set to 1. The
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    TaKaRa mir x mirna qrt pcr sybr kit
    IL-4 is a direct target of miR-340/429 A . IL-4 targeting-eight miRNAs were selected using <t>miRNA</t> prediction sites and were confirmed by Western blot analysis (left). IR-inhibited 26 miRNAs were picked according to the previous study of Weidhaas and Chaudhry group [ 36 , 37 ] (right). miR-340 and miR-429 were selected as IR-inhibited and IL-4-targeting miRNAs for further investigation. B . After a single or fractionated IR exposure to A498 cells (5 Gy) (left) or mouse (2.5Gy x 3 times) (right) respectively, the expression of miR-340 or miR-429 in cells or mouse pulmonary tissues was quantified by <t>qRT-PCR.</t> The data are presented as the mean S.D. (** P
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    Thermo Fisher cellsdirect one step qrt pcr kit
    HTM cell characterization. ( A ) Representative phase contrast micrographs of reference, HTM01, HTM11, and HTM12 cell strains with sex/age information. Scale bar, 250 µm. ( B ) mRNA fold-change of MYOC by <t>qRT-PCR</t> at 7 d (N = 3 per group and donor; *p
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    TaKaRa lenti x qrt pcr titration kit
    Transduction and RNAi efficiency of two human Nav1.1 lenti-shRNA clones. ( A ) Evaluating the transduction efficiency of the lentiviral particles were tested in HEK293T cells based on the expression of the tRFP reporter. Scale bars, 20 microns. ( B ) The RNAi efficiency of lenti-shRNA1 and lenti-shRNA2 relative to a non-silencing clone was determined by first transducing HEK293T cells with the lentiviral clones and next expressing the human Na v 1.1 cDNA via transfection. Na v 1.1 mRNAs were quantified with <t>qRT-PCR.</t> By ANOVA and post hoc Sidak’s multiple comparisons, p = 0.003 for the comparison between control and shRNA1; p = 0.002 for the comparison between control and shRNA2. DOI: http://dx.doi.org/10.7554/eLife.13073.034
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    Image Search Results


    miR-155 promotes production of TNF-α and IL-1β in human PBMCs. ( A ) Fold changes of miR-155 expression were determined by qRT-PCR after the transfection of miR-control, or miR-155 mimics at 25 or 50 nM for 24 h; ( B ) and ( C ) Expression of TNF-α and IL-1β in supernatant of PBMCs 24 h post LPS treatment, or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM miRNA control. Results were expressed as pg/mL by an ELISA test; and ( D ) mRNA expression of TNF-α (group 1) and IL-1β (group 2) in PBMCs 24 h after LPS treatment or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM miRNA control. All results are the average of three independent experiments. Statistical significance is shown.

    Journal: International Journal of Molecular Sciences

    Article Title: Rheumatoid Arthritis-Associated MicroRNA-155 Targets SOCS1 and Upregulates TNF-? and IL-1? in PBMCs

    doi: 10.3390/ijms141223910

    Figure Lengend Snippet: miR-155 promotes production of TNF-α and IL-1β in human PBMCs. ( A ) Fold changes of miR-155 expression were determined by qRT-PCR after the transfection of miR-control, or miR-155 mimics at 25 or 50 nM for 24 h; ( B ) and ( C ) Expression of TNF-α and IL-1β in supernatant of PBMCs 24 h post LPS treatment, or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM miRNA control. Results were expressed as pg/mL by an ELISA test; and ( D ) mRNA expression of TNF-α (group 1) and IL-1β (group 2) in PBMCs 24 h after LPS treatment or 24 h post transfection with 25 nM miR-155 mimics, 50 nM miR-155 mimics or 50 nM miRNA control. All results are the average of three independent experiments. Statistical significance is shown.

    Article Snippet: Quantification of miR-155 expression was conducted using the mirVana qRT-PCR miRNA Detection Kit (Ambion, Austin, TX, USA), and the U6 small nuclear RNA was used as internal control.

    Techniques: Expressing, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay

    After osteogenic induction, total RNA was extracted from all groups and expression of RUNX 2 and OCN was assessed by qRT-PCR. The expression of RUNX 2 and OCN of hUSCs (P4) from the children group was higher than that from the elder group (* P

    Journal: Cytotechnology

    Article Title: Effects of the donor age on proliferation, senescence and osteogenic capacity of human urine-derived stem cells

    doi: 10.1007/s10616-017-0084-5

    Figure Lengend Snippet: After osteogenic induction, total RNA was extracted from all groups and expression of RUNX 2 and OCN was assessed by qRT-PCR. The expression of RUNX 2 and OCN of hUSCs (P4) from the children group was higher than that from the elder group (* P

    Article Snippet: The mRNA levels of RUNX2 , OCN and β - actin were determined by quantitative real-time PCR (qRT-PCR) using C1000 instrument (Bio-Rad, Hercules, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR

    Gestational stress induces differential miRNA expression in frontal cortex. A, Schematic overview of miRNA biogenesis pathways. B, Heat map representation of differentially regulated miRNAs, as observed by microarray analysis. C, Table of target genes for miRNAs modulated by gestational stress (miR-329, miR-380, miR-20a, and miR-500; p≤0.05), and their physiological implications. D, Expression ratio group averages of miRNAs as observed by qRT-PCR analysis (p≤0.05). Note that prenatal stress downregulated miR-181 and miR-186 expression in the frontal cortex. miRNA analyses were performed in dams that showed representative behavioural characteristics (n = 3 per group, three repeats per sample). All data are presented as mean ± SEM.

    Journal: PLoS ONE

    Article Title: Maternal Stress Induces Epigenetic Signatures of Psychiatric and Neurological Diseases in the Offspring

    doi: 10.1371/journal.pone.0056967

    Figure Lengend Snippet: Gestational stress induces differential miRNA expression in frontal cortex. A, Schematic overview of miRNA biogenesis pathways. B, Heat map representation of differentially regulated miRNAs, as observed by microarray analysis. C, Table of target genes for miRNAs modulated by gestational stress (miR-329, miR-380, miR-20a, and miR-500; p≤0.05), and their physiological implications. D, Expression ratio group averages of miRNAs as observed by qRT-PCR analysis (p≤0.05). Note that prenatal stress downregulated miR-181 and miR-186 expression in the frontal cortex. miRNA analyses were performed in dams that showed representative behavioural characteristics (n = 3 per group, three repeats per sample). All data are presented as mean ± SEM.

    Article Snippet: The generation of cDNAs from the total RNA samples was performed using M-MuLV Reverse Transcriptase, NEB#M0253S (New England Biolab, Ipswich, MA; see for RT primers). qRT-PCR reactions were conducted with Bio-Rad CFX96™ Real-Time PCR Systems, using SsoFast™ EvaGreen® Supermix (Bio-Rad, Mississauga, ON) reaction premix added to the cDNAs templates and specific primers, according to the manufacturer’s protocol (see for primer reference).

    Techniques: Expressing, Microarray, Quantitative RT-PCR

    Prenatal stress modulates the brain miRNAome in male newborn offspring. A, Heat map representation of differentially regulated miRNA as observed by microarray analyses. B, Table of putative target genes for modulated miRNAs (miR-103, miR-151, and miR-219-2-3p; p≤0.05) and their physiological functions. C, Expression ratio group averages of miRNAs as observed by qRT-PCR analysis (p≤0.05). Whole brains of newborns born to dams shown in Figures 1 and 2 (n = 3 per group, three repeats per sample; 1 pup per dam) were used. All data are presented as mean ± SEM.

    Journal: PLoS ONE

    Article Title: Maternal Stress Induces Epigenetic Signatures of Psychiatric and Neurological Diseases in the Offspring

    doi: 10.1371/journal.pone.0056967

    Figure Lengend Snippet: Prenatal stress modulates the brain miRNAome in male newborn offspring. A, Heat map representation of differentially regulated miRNA as observed by microarray analyses. B, Table of putative target genes for modulated miRNAs (miR-103, miR-151, and miR-219-2-3p; p≤0.05) and their physiological functions. C, Expression ratio group averages of miRNAs as observed by qRT-PCR analysis (p≤0.05). Whole brains of newborns born to dams shown in Figures 1 and 2 (n = 3 per group, three repeats per sample; 1 pup per dam) were used. All data are presented as mean ± SEM.

    Article Snippet: The generation of cDNAs from the total RNA samples was performed using M-MuLV Reverse Transcriptase, NEB#M0253S (New England Biolab, Ipswich, MA; see for RT primers). qRT-PCR reactions were conducted with Bio-Rad CFX96™ Real-Time PCR Systems, using SsoFast™ EvaGreen® Supermix (Bio-Rad, Mississauga, ON) reaction premix added to the cDNAs templates and specific primers, according to the manufacturer’s protocol (see for primer reference).

    Techniques: Microarray, Expressing, Quantitative RT-PCR

    EOMES expression is increased in D*V embryonic comb tissue. Results of qRT-PCR analysis demonstrating increased EOMES expression in D*V embryonic comb tissue whereas CMC1 and AZI2 , located nearby the duplicated region, do not show any significant change in expression. Bar graphs mean ±sem, ANOVA, *P

    Journal: PLoS Genetics

    Article Title: A Genomic Duplication is Associated with Ectopic Eomesodermin Expression in the Embryonic Chicken Comb and Two Duplex-comb Phenotypes

    doi: 10.1371/journal.pgen.1004947

    Figure Lengend Snippet: EOMES expression is increased in D*V embryonic comb tissue. Results of qRT-PCR analysis demonstrating increased EOMES expression in D*V embryonic comb tissue whereas CMC1 and AZI2 , located nearby the duplicated region, do not show any significant change in expression. Bar graphs mean ±sem, ANOVA, *P

    Article Snippet: The qRT-PCR analysis was performed using CFX96 SyBr Green Supermix (Bio-Rad) with primers designed by using Primer Express v2.0 (ABI), checked for PCR efficiency, linear dynamic range and specificity.

    Techniques: Expressing, Quantitative RT-PCR

    Inhibition of SF3B1 reduces HIV products. (A) U87/CD4/CXCR4 cells were infected with replication-competent HIV-1 for 24 h. Total cellular RNA was obtained, and qRT-PCR was performed for the unspliced (US), singly spliced (SS), and multiply spliced (MS) forms of HIV using specific primers. RNA fold change was calculated relative to DMSO treatment. (B) 293T cells were infected with HIV Δ env for 24 h in the presence of DMSO or sudemycin D6. Western blots were performed on cell lysates for HIV Gag products. Three experiments were quantitated (C). (D) 293T cells were infected with HIV Δ env for 24 h in the presence of DMSO or sudemycin D6. Western blots were performed on cell lysates for Tat and Rev. Data indicate means, and error bars indicate ± SEM ( n = 3). **, P

    Journal: mBio

    Article Title: Splicing Factor 3B Subunit 1 Interacts with HIV Tat and Plays a Role in Viral Transcription and Reactivation from Latency

    doi: 10.1128/mBio.01423-18

    Figure Lengend Snippet: Inhibition of SF3B1 reduces HIV products. (A) U87/CD4/CXCR4 cells were infected with replication-competent HIV-1 for 24 h. Total cellular RNA was obtained, and qRT-PCR was performed for the unspliced (US), singly spliced (SS), and multiply spliced (MS) forms of HIV using specific primers. RNA fold change was calculated relative to DMSO treatment. (B) 293T cells were infected with HIV Δ env for 24 h in the presence of DMSO or sudemycin D6. Western blots were performed on cell lysates for HIV Gag products. Three experiments were quantitated (C). (D) 293T cells were infected with HIV Δ env for 24 h in the presence of DMSO or sudemycin D6. Western blots were performed on cell lysates for Tat and Rev. Data indicate means, and error bars indicate ± SEM ( n = 3). **, P

    Article Snippet: Real-time qRT-PCR was done with the Bio-Rad One-Step RT-PCR kit using the Bio-Rad CFX Connect system with SYBR green.

    Techniques: Inhibition, Infection, Quantitative RT-PCR, Mass Spectrometry, Western Blot

    Inhibition of SF3B1 prevents HIV reactivation from latency. (A) JLAT10.6 cells were incubated with the indicated compounds for 24 h, and FACS was performed to detect live GFP-positive cells as a measure of HIV reactivation from latency. (B) JLAT10.6 cells were incubated with the indicated compounds for 24 h. Cells were lysed and resolved on Western blots for the HIV products. A representative blot of 3 independent experiments is shown. (C) qRT-PCR for a similar experiment in panel B. Fold change is relative to DMSO treatment. (D) Inhibition of HIV reactivation in the Greene model of latency. Resting CD4 + T cells were infected with HIV-1 Luc, treated with protease inhibitor darunavir for 2 days. Cells were incubated with the compounds shown for 24 h in the presence of raltegravir. (See Materials and Methods for details.) HIV reactivation was measured by luciferase-based luminescence in the cell lysates normalized to protein concentration. (E) JLAT10.6 cells were treated with 5 μM sudemycin D6 for 18 h. Cells were washed and allowed to recover for 24 h. At 24, 48, or 72 h after drug washout, the cells were treated with LRAs for 24 h. HIV reactivation was measured with FACS to detect live cells expressing GFP. Data indicate means, and error bars indicate ±SEM ( n = 3). **, P

    Journal: mBio

    Article Title: Splicing Factor 3B Subunit 1 Interacts with HIV Tat and Plays a Role in Viral Transcription and Reactivation from Latency

    doi: 10.1128/mBio.01423-18

    Figure Lengend Snippet: Inhibition of SF3B1 prevents HIV reactivation from latency. (A) JLAT10.6 cells were incubated with the indicated compounds for 24 h, and FACS was performed to detect live GFP-positive cells as a measure of HIV reactivation from latency. (B) JLAT10.6 cells were incubated with the indicated compounds for 24 h. Cells were lysed and resolved on Western blots for the HIV products. A representative blot of 3 independent experiments is shown. (C) qRT-PCR for a similar experiment in panel B. Fold change is relative to DMSO treatment. (D) Inhibition of HIV reactivation in the Greene model of latency. Resting CD4 + T cells were infected with HIV-1 Luc, treated with protease inhibitor darunavir for 2 days. Cells were incubated with the compounds shown for 24 h in the presence of raltegravir. (See Materials and Methods for details.) HIV reactivation was measured by luciferase-based luminescence in the cell lysates normalized to protein concentration. (E) JLAT10.6 cells were treated with 5 μM sudemycin D6 for 18 h. Cells were washed and allowed to recover for 24 h. At 24, 48, or 72 h after drug washout, the cells were treated with LRAs for 24 h. HIV reactivation was measured with FACS to detect live cells expressing GFP. Data indicate means, and error bars indicate ±SEM ( n = 3). **, P

    Article Snippet: Real-time qRT-PCR was done with the Bio-Rad One-Step RT-PCR kit using the Bio-Rad CFX Connect system with SYBR green.

    Techniques: Inhibition, Incubation, FACS, Western Blot, Quantitative RT-PCR, Infection, Protease Inhibitor, Luciferase, Protein Concentration, Expressing

    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using qRT-PCRs. (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.

    Journal: Scientific Reports

    Article Title: Genome-wide identification of Arabidopsis long noncoding RNAs in response to the blue light

    doi: 10.1038/s41598-020-63187-1

    Figure Lengend Snippet: The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using qRT-PCRs. (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.

    Article Snippet: The primers used for qRT-PCRs are listed in Table S2 supplemental information.

    Techniques: Expressing, Northern Blot

    Effect of NLRC5 KD on differentially expressed immune genes during H5N2 infection. NLRC5 siRNA and scramble control transfected MQ-NCSU cells were infected with H5N2 virus. Total RNA was collected at 24 hpi and assayed by the RT 2 Profiler PCR Array system. Selected genes taking part in innate and adaptive immune response are shown as log 2 transform of fold-change expression relative to the gene expression in the mock-transfected cells. Data represent an average of three biological replicates.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: NLRC5 Serves as a Pro-viral Factor During Influenza Virus Infection in Chicken Macrophages

    doi: 10.3389/fcimb.2020.00230

    Figure Lengend Snippet: Effect of NLRC5 KD on differentially expressed immune genes during H5N2 infection. NLRC5 siRNA and scramble control transfected MQ-NCSU cells were infected with H5N2 virus. Total RNA was collected at 24 hpi and assayed by the RT 2 Profiler PCR Array system. Selected genes taking part in innate and adaptive immune response are shown as log 2 transform of fold-change expression relative to the gene expression in the mock-transfected cells. Data represent an average of three biological replicates.

    Article Snippet: The viral matrix (M) gene was amplified with the SuperScript™ III Platinum™ One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA, USA) using primers and Taqman probe previously described (Spackman et al., ).

    Techniques: Infection, Transfection, Polymerase Chain Reaction, Expressing

    Reduced virus titers of LPAIV and HPAIV in NLRC5 KD chicken macrophages. MQ-NCSU cells were transfected with NLRC5 siRNA or non-specific control siRNA (scramble) and infected with H5N2, H7N2 or H5N1 at 32 hpt. NLRC5 mRNA and viral RNA were quantified by qRT-PCR. (A) NLRC5 mRNA levels were measured at 32 hpt and compared to control. Viral RNA levels were measured by M-gene amplification at 8 and 24 hpi with (B) H5N2 and (C) H5N1. Supernatant infectious virus production was measured using relative equivalence units (REUs) at 8 and 24 hpi with (D) H5N2 or (E) H7N2. Data represents average of three biological replicates with error bars showing standard deviation. * p ≤ 0.05, ** p ≤ 0.01.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: NLRC5 Serves as a Pro-viral Factor During Influenza Virus Infection in Chicken Macrophages

    doi: 10.3389/fcimb.2020.00230

    Figure Lengend Snippet: Reduced virus titers of LPAIV and HPAIV in NLRC5 KD chicken macrophages. MQ-NCSU cells were transfected with NLRC5 siRNA or non-specific control siRNA (scramble) and infected with H5N2, H7N2 or H5N1 at 32 hpt. NLRC5 mRNA and viral RNA were quantified by qRT-PCR. (A) NLRC5 mRNA levels were measured at 32 hpt and compared to control. Viral RNA levels were measured by M-gene amplification at 8 and 24 hpi with (B) H5N2 and (C) H5N1. Supernatant infectious virus production was measured using relative equivalence units (REUs) at 8 and 24 hpi with (D) H5N2 or (E) H7N2. Data represents average of three biological replicates with error bars showing standard deviation. * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: The viral matrix (M) gene was amplified with the SuperScript™ III Platinum™ One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA, USA) using primers and Taqman probe previously described (Spackman et al., ).

    Techniques: Transfection, Infection, Quantitative RT-PCR, Amplification, Standard Deviation

    Reduced mRNA expression of IL-1β and IL-18 in IAV infected NLRC5 KD cells. MQ-NCSU cells were transfected with NLRC5 siRNA or non-specific control siRNA (scramble). The relative mRNA expression of IL-1β and IL-18 was determined at 32 hpt by quantitative RT-PCR and normalized to the housekeeping gene HMBS. Data represents average of three biological replicates with error bars showing standard deviation. * p ≤ 0.05, ** p ≤ 0.01.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: NLRC5 Serves as a Pro-viral Factor During Influenza Virus Infection in Chicken Macrophages

    doi: 10.3389/fcimb.2020.00230

    Figure Lengend Snippet: Reduced mRNA expression of IL-1β and IL-18 in IAV infected NLRC5 KD cells. MQ-NCSU cells were transfected with NLRC5 siRNA or non-specific control siRNA (scramble). The relative mRNA expression of IL-1β and IL-18 was determined at 32 hpt by quantitative RT-PCR and normalized to the housekeeping gene HMBS. Data represents average of three biological replicates with error bars showing standard deviation. * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: The viral matrix (M) gene was amplified with the SuperScript™ III Platinum™ One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA, USA) using primers and Taqman probe previously described (Spackman et al., ).

    Techniques: Expressing, Infection, Transfection, Quantitative RT-PCR, Standard Deviation

    In vitro and ex vivo transcriptional regulation of chIFIT5 gene. ( A ) ChIFN-β itself, or produced by different stimuli, can initiate JAK-STAT signalling pathway and culminate in the induction of hundreds of ISGs. IFIT family of genes is one such antiviral effector of innate immune responses. ( B ) Quantitation of chIFIT5 mRNA in cells stimulated with 1000 U of chIFN-β, 10 μg/mL of LPS, 5 μg/mL of poly I:C or 1 MOI of NDV for 24 hours before RNA extraction and analysis for qRT-PCR using primers specific for the chIFIT5 gene. ( C–F ) Total cellular RNA was extracted from CEF cells at 1, 2, 4, 8, 16, and 24 hours post-NDV infection and was subjected to quantitation of mRNA for chIFIT5 ( C ), viral mRNA ( D ), chMx ( E ) and chIFN-β ( F , G ) RNA collected from seven organs infected or not with H9N2 influenza viruses were used to determine the level of chIFIT5 mRNA and M gene of the virus. Fold change induction in all experiments was determined by 2 −∆∆CT algorithm and data presented is average of three independent experiments.

    Journal: Scientific Reports

    Article Title: Chicken Interferon-induced Protein with Tetratricopeptide Repeats 5 Antagonizes Replication of RNA Viruses

    doi: 10.1038/s41598-018-24905-y

    Figure Lengend Snippet: In vitro and ex vivo transcriptional regulation of chIFIT5 gene. ( A ) ChIFN-β itself, or produced by different stimuli, can initiate JAK-STAT signalling pathway and culminate in the induction of hundreds of ISGs. IFIT family of genes is one such antiviral effector of innate immune responses. ( B ) Quantitation of chIFIT5 mRNA in cells stimulated with 1000 U of chIFN-β, 10 μg/mL of LPS, 5 μg/mL of poly I:C or 1 MOI of NDV for 24 hours before RNA extraction and analysis for qRT-PCR using primers specific for the chIFIT5 gene. ( C–F ) Total cellular RNA was extracted from CEF cells at 1, 2, 4, 8, 16, and 24 hours post-NDV infection and was subjected to quantitation of mRNA for chIFIT5 ( C ), viral mRNA ( D ), chMx ( E ) and chIFN-β ( F , G ) RNA collected from seven organs infected or not with H9N2 influenza viruses were used to determine the level of chIFIT5 mRNA and M gene of the virus. Fold change induction in all experiments was determined by 2 −∆∆CT algorithm and data presented is average of three independent experiments.

    Article Snippet: A total of 200 ng of RNA was used in PCR reactions using SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA, USA).

    Techniques: In Vitro, Ex Vivo, Produced, Quantitation Assay, RNA Extraction, Quantitative RT-PCR, Infection

    Silencing of endogenous chIFIT5 gene in transgenic chicken embryos and impact on the virus replication. ( A ) Three shRNA targeting exon 2 of chIFIT5 were cloned in pRFPRNAiC vector between NheI and MluI downstream to the chicken U6 promoter. ( B ) DF-1 cells were transfected with 500 ng of pRFPRNAiC-shRNA plasmid expressing each of the cloned shRNA or empty vector. The expression of RFP marker gene demonstrates the integrity of the constructs. ( C ) Transfer of validated shRNA (shRNA #3) cassette between NotI and ClaI sites in RCASBP(A) vector. ( D ) Silencing efficacies of all three chIFIT5-targeted shRNA compared to scrambled (non-targeting) shRNA. DF-1 cells, transfected with 500 ng of each of the plasmids, were used to extract total cellular RNA and the level of chIFIT5 gene silencing was monitored using qRT-PCR. ( E ) Retroviruses were rescued in DF-1 cells and stained for structural gag protein indicating the replication-competency of these retroviruses. ( F ) Infectious cells stably expressing pre-validated shRNA against chIFIT5 gene were inoculated in 3 days old chicken embryos and allowed to develop until 9 days post-embryonation when 100 FPUs of NDV were inoculated per transgenic egg and the quantification of the virus replication was performed on day 14 post-embryonation. ( G ) Quantitative analysis of viruses in NDV-infected and un-infected transgenic embryos expressing shRNA, wt chIFIT5 or shRNA-resistant chIFIT5. Each dot represents individual chicken embryo in all experimental groups.

    Journal: Scientific Reports

    Article Title: Chicken Interferon-induced Protein with Tetratricopeptide Repeats 5 Antagonizes Replication of RNA Viruses

    doi: 10.1038/s41598-018-24905-y

    Figure Lengend Snippet: Silencing of endogenous chIFIT5 gene in transgenic chicken embryos and impact on the virus replication. ( A ) Three shRNA targeting exon 2 of chIFIT5 were cloned in pRFPRNAiC vector between NheI and MluI downstream to the chicken U6 promoter. ( B ) DF-1 cells were transfected with 500 ng of pRFPRNAiC-shRNA plasmid expressing each of the cloned shRNA or empty vector. The expression of RFP marker gene demonstrates the integrity of the constructs. ( C ) Transfer of validated shRNA (shRNA #3) cassette between NotI and ClaI sites in RCASBP(A) vector. ( D ) Silencing efficacies of all three chIFIT5-targeted shRNA compared to scrambled (non-targeting) shRNA. DF-1 cells, transfected with 500 ng of each of the plasmids, were used to extract total cellular RNA and the level of chIFIT5 gene silencing was monitored using qRT-PCR. ( E ) Retroviruses were rescued in DF-1 cells and stained for structural gag protein indicating the replication-competency of these retroviruses. ( F ) Infectious cells stably expressing pre-validated shRNA against chIFIT5 gene were inoculated in 3 days old chicken embryos and allowed to develop until 9 days post-embryonation when 100 FPUs of NDV were inoculated per transgenic egg and the quantification of the virus replication was performed on day 14 post-embryonation. ( G ) Quantitative analysis of viruses in NDV-infected and un-infected transgenic embryos expressing shRNA, wt chIFIT5 or shRNA-resistant chIFIT5. Each dot represents individual chicken embryo in all experimental groups.

    Article Snippet: A total of 200 ng of RNA was used in PCR reactions using SuperScript® III Platinum® SYBR® Green One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA, USA).

    Techniques: Transgenic Assay, shRNA, Clone Assay, Plasmid Preparation, Transfection, Expressing, Marker, Construct, Quantitative RT-PCR, Staining, Stable Transfection, Infection

    TAp63 modulates the expression of miR-133b. HT-29 and SW-620 cells were transfected with either an empty vector or with TAp63 for 72 h. ( A ) Endogenous TAp63 levels were assessed by RT–PCR. ( B ) The expression level of TAp63 is shown in the western blotting. ( C ). Endogenous TAp63 levels were again confirmed by qRT–PCR. As the figure shows, HT-29 and SW-620 cells transfected with TAp63 expressing vector induced a significant increase in TAp63 expression. ( D ) Endogenous miR-133b levels were assessed by qRT–PCR. After transfection with a TAp63-expressing vector, HT-29 and SW-620 cells showed a significant increase in miR-133b expression. ( E ) Chromatin immunoprecipitation (ChIP) experiment showed that p63 is able to bind the p53 RE-III site but not p53 RE-I and p53 RE-II. ( F ) Insertion of miR-133b promoter region in a luciferase reporter gene leads to increased luciferase activity in the presence of TAp63 in HT-29 and SW-620 cells. Mutation of the RE-III p53-binding site abolished TAp63-mediated luciferase activity. ( G ) The miR-133b promoter region containing the three p53 consensus sites (p53RE). All qRT–PCR results were relative to U6-snRNA or GAPDH and were normalised to the expression of miR-133b or TAp63 in HT-29 cells. Data represent the means±s.d. of three different experiments analysed in triplicate. * P

    Journal: British Journal of Cancer

    Article Title: TAp63 suppress metastasis via miR-133b in colon cancer cells

    doi: 10.1038/bjc.2014.118

    Figure Lengend Snippet: TAp63 modulates the expression of miR-133b. HT-29 and SW-620 cells were transfected with either an empty vector or with TAp63 for 72 h. ( A ) Endogenous TAp63 levels were assessed by RT–PCR. ( B ) The expression level of TAp63 is shown in the western blotting. ( C ). Endogenous TAp63 levels were again confirmed by qRT–PCR. As the figure shows, HT-29 and SW-620 cells transfected with TAp63 expressing vector induced a significant increase in TAp63 expression. ( D ) Endogenous miR-133b levels were assessed by qRT–PCR. After transfection with a TAp63-expressing vector, HT-29 and SW-620 cells showed a significant increase in miR-133b expression. ( E ) Chromatin immunoprecipitation (ChIP) experiment showed that p63 is able to bind the p53 RE-III site but not p53 RE-I and p53 RE-II. ( F ) Insertion of miR-133b promoter region in a luciferase reporter gene leads to increased luciferase activity in the presence of TAp63 in HT-29 and SW-620 cells. Mutation of the RE-III p53-binding site abolished TAp63-mediated luciferase activity. ( G ) The miR-133b promoter region containing the three p53 consensus sites (p53RE). All qRT–PCR results were relative to U6-snRNA or GAPDH and were normalised to the expression of miR-133b or TAp63 in HT-29 cells. Data represent the means±s.d. of three different experiments analysed in triplicate. * P

    Article Snippet: MiRNA reverse transcription was performed using an All-in-One miRNA qRT–PCR Detection Kit (GeneCopoeia, Rockville, MD, USA).

    Techniques: Expressing, Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Chromatin Immunoprecipitation, Luciferase, Activity Assay, Mutagenesis, Binding Assay

    Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by qRT-PCR. (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of three biological replicates. ** P

    Journal: Emerging Microbes & Infections

    Article Title: Rsp promotes the transcription of virulence factors in an agr-independent manner in Staphylococcus aureus

    doi: 10.1080/22221751.2020.1752116

    Figure Lengend Snippet: Rsp positively regulates its own expression. (A) The transcript levels of rsp at different growth phases were measured by qRT-PCR. (B) The β-galactosidase activities of the WT and rsp mutant strains with a plasmid pOS rsp were detected in the indicated time points. (C) Analysis of mRNA half-lives of p rsp - lacZ and rsp . Cells were collected after rifampin (200 μg/ml) treatment for RNA isolation, and then the mRNA half-lives were measured by qRT-PCR. (D) SDS-PAGE analysis of Rsp purified from the pRSF-Duet expression vector. The gel was stained with Coomassie blue. (E) EMSA of purified Rsp with the biotin-labelled DNA fragment containing the putative promoter region of rsp . Increasing concentrations of purified Rsp and 4 fmol of the biotin-labelled probe were used in the reactions. The unlabelled probe was added as a specific competitor, and the unlabelled fragment of the hu ORF region was added as a nonspecific competitor. (F) Promoter sequence of the rsp gene. The start points of the truncated probes are marked by arrows. (G) EMSA assay of Rsp with rsp truncated probes. The nonspecific competitor was added to the reactions. The error bars indicate the standard errors of the means of three biological replicates. ** P

    Article Snippet: Total RNA isolation, cDNA generation, and real-time quantitative reverse transcription-PCR Overnight cultures of S. aureus were diluted 1:100 in TSB and then grown to the indicated cell density until being collected.

    Techniques: Expressing, Quantitative RT-PCR, Mutagenesis, Plasmid Preparation, Isolation, SDS Page, Purification, Staining, Sequencing

    Rsp positively regulates the transcription of virulence genes. (A) The transcript levels of 7 virulence-related genes were detected by qRT-PCR in the WT and rs p mutant strains. The transcript levels of hla (B), psmα (C), and psmβ (D) in the WT, rs p mutant, and rs p chromosomal-complemented strains at different growth phases. The β-galactosidase activities of hla (E), psmα (F), and psmβ (G) promoter in the WT and rs p mutant strains. Cells were collected at early stationary phase, and the β-galactosidase activity was detected with ONPG. (H) Hla protein levels of the WT, rs p mutant, and rs p chromosomal-complemented strains were measured by Western blot. (I) The transcript levels of hla , psmα , and psmβ in the WT, rs p mutant, and rs p chromosomal-complemented strains in the process of haemolysis. Cells were collected after incubated with 3% sheep red blood cells for 30 min, and the mRNA levels were analysed by qRT-PCR. The error bars indicate the standard errors of the means of three biological replicates. * P

    Journal: Emerging Microbes & Infections

    Article Title: Rsp promotes the transcription of virulence factors in an agr-independent manner in Staphylococcus aureus

    doi: 10.1080/22221751.2020.1752116

    Figure Lengend Snippet: Rsp positively regulates the transcription of virulence genes. (A) The transcript levels of 7 virulence-related genes were detected by qRT-PCR in the WT and rs p mutant strains. The transcript levels of hla (B), psmα (C), and psmβ (D) in the WT, rs p mutant, and rs p chromosomal-complemented strains at different growth phases. The β-galactosidase activities of hla (E), psmα (F), and psmβ (G) promoter in the WT and rs p mutant strains. Cells were collected at early stationary phase, and the β-galactosidase activity was detected with ONPG. (H) Hla protein levels of the WT, rs p mutant, and rs p chromosomal-complemented strains were measured by Western blot. (I) The transcript levels of hla , psmα , and psmβ in the WT, rs p mutant, and rs p chromosomal-complemented strains in the process of haemolysis. Cells were collected after incubated with 3% sheep red blood cells for 30 min, and the mRNA levels were analysed by qRT-PCR. The error bars indicate the standard errors of the means of three biological replicates. * P

    Article Snippet: Total RNA isolation, cDNA generation, and real-time quantitative reverse transcription-PCR Overnight cultures of S. aureus were diluted 1:100 in TSB and then grown to the indicated cell density until being collected.

    Techniques: Quantitative RT-PCR, Mutagenesis, Activity Assay, Western Blot, Incubation

    miR-370 regulates MGMT expression in raji cells. a qRT-PCR demonstrated that miR-370 inhibitors increased expression of MGMT at mRNA level, while miR-370 mimics suppressed MGMT mRNA expression. b The result of MGMT protein level examined by western blot was similar with ( a ) after being transfected with miR-370 inhibitors or mimics

    Journal: Pathology Oncology Research

    Article Title: miR-370 Sensitizes TMZ Response Dependent of MGMT Status in Primary Central Nervous System Lymphoma

    doi: 10.1007/s12253-019-00605-4

    Figure Lengend Snippet: miR-370 regulates MGMT expression in raji cells. a qRT-PCR demonstrated that miR-370 inhibitors increased expression of MGMT at mRNA level, while miR-370 mimics suppressed MGMT mRNA expression. b The result of MGMT protein level examined by western blot was similar with ( a ) after being transfected with miR-370 inhibitors or mimics

    Article Snippet: The process quantitative of real-time polymerase chain reaction (qRT-PCR) was performed according to the manufacturer’s instructions [ ].

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection

    Expression of MGMT and miR-370 were detected in PCNSL and nodal tissues. a Immunohistochemistry revealed high expression of MGMT in twenty clinical specimens. b qRT-PCR results showed that miR-370 was differentially downregulated in PCNSL tissues compared to nodal tissues

    Journal: Pathology Oncology Research

    Article Title: miR-370 Sensitizes TMZ Response Dependent of MGMT Status in Primary Central Nervous System Lymphoma

    doi: 10.1007/s12253-019-00605-4

    Figure Lengend Snippet: Expression of MGMT and miR-370 were detected in PCNSL and nodal tissues. a Immunohistochemistry revealed high expression of MGMT in twenty clinical specimens. b qRT-PCR results showed that miR-370 was differentially downregulated in PCNSL tissues compared to nodal tissues

    Article Snippet: The process quantitative of real-time polymerase chain reaction (qRT-PCR) was performed according to the manufacturer’s instructions [ ].

    Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR

    xnp1 gene expression is induced by mitomycin C treatment. Results shown are for qRT-PCR analysis of xnp1 gene expression before treatment with mitomycin C (0 h) and 1 h and 2.5 h after induction. For each gene, time zero was arbitrarily set to 1. The

    Journal: Journal of Bacteriology

    Article Title: The xnp1 P2-Like Tail Synthesis Gene Cluster Encodes Xenorhabdicin and Is Required for Interspecies Competition ▿ P2-Like Tail Synthesis Gene Cluster Encodes Xenorhabdicin and Is Required for Interspecies Competition ▿ †

    doi: 10.1128/JB.00092-11

    Figure Lengend Snippet: xnp1 gene expression is induced by mitomycin C treatment. Results shown are for qRT-PCR analysis of xnp1 gene expression before treatment with mitomycin C (0 h) and 1 h and 2.5 h after induction. For each gene, time zero was arbitrarily set to 1. The

    Article Snippet: For negative-control experiments, reactions were conducted without reverse transcriptase. qRT-PCR was performed with the SuperScript III Platinum two-step qRT-PCR kit with SYBR green (Invitrogen).

    Techniques: Expressing, Quantitative RT-PCR

    IL-4 is a direct target of miR-340/429 A . IL-4 targeting-eight miRNAs were selected using miRNA prediction sites and were confirmed by Western blot analysis (left). IR-inhibited 26 miRNAs were picked according to the previous study of Weidhaas and Chaudhry group [ 36 , 37 ] (right). miR-340 and miR-429 were selected as IR-inhibited and IL-4-targeting miRNAs for further investigation. B . After a single or fractionated IR exposure to A498 cells (5 Gy) (left) or mouse (2.5Gy x 3 times) (right) respectively, the expression of miR-340 or miR-429 in cells or mouse pulmonary tissues was quantified by qRT-PCR. The data are presented as the mean S.D. (** P

    Journal: Oncotarget

    Article Title: IL-4, a direct target of miR-340/429, is involved in radiation-induced aggressive tumor behavior in human carcinoma cells

    doi: 10.18632/oncotarget.13561

    Figure Lengend Snippet: IL-4 is a direct target of miR-340/429 A . IL-4 targeting-eight miRNAs were selected using miRNA prediction sites and were confirmed by Western blot analysis (left). IR-inhibited 26 miRNAs were picked according to the previous study of Weidhaas and Chaudhry group [ 36 , 37 ] (right). miR-340 and miR-429 were selected as IR-inhibited and IL-4-targeting miRNAs for further investigation. B . After a single or fractionated IR exposure to A498 cells (5 Gy) (left) or mouse (2.5Gy x 3 times) (right) respectively, the expression of miR-340 or miR-429 in cells or mouse pulmonary tissues was quantified by qRT-PCR. The data are presented as the mean S.D. (** P

    Article Snippet: The level of miR-340 or -429 were quantified by real-time qRT-PCR using Mir-X miRNA qRT-PCR SYBR kit (Clontech Laboratories Inc., Mountain view, CA, USA).

    Techniques: Western Blot, Expressing, Quantitative RT-PCR

    HTM cell characterization. ( A ) Representative phase contrast micrographs of reference, HTM01, HTM11, and HTM12 cell strains with sex/age information. Scale bar, 250 µm. ( B ) mRNA fold-change of MYOC by qRT-PCR at 7 d (N = 3 per group and donor; *p

    Journal: bioRxiv

    Article Title: A tissue-engineered human trabecular meshwork hydrogel for advanced glaucoma disease modeling

    doi: 10.1101/2020.07.31.229229

    Figure Lengend Snippet: HTM cell characterization. ( A ) Representative phase contrast micrographs of reference, HTM01, HTM11, and HTM12 cell strains with sex/age information. Scale bar, 250 µm. ( B ) mRNA fold-change of MYOC by qRT-PCR at 7 d (N = 3 per group and donor; *p

    Article Snippet: The monolayer HTM cells were processed for quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunocytochemistry (ICC) analyses.

    Techniques: Quantitative RT-PCR

    Transduction and RNAi efficiency of two human Nav1.1 lenti-shRNA clones. ( A ) Evaluating the transduction efficiency of the lentiviral particles were tested in HEK293T cells based on the expression of the tRFP reporter. Scale bars, 20 microns. ( B ) The RNAi efficiency of lenti-shRNA1 and lenti-shRNA2 relative to a non-silencing clone was determined by first transducing HEK293T cells with the lentiviral clones and next expressing the human Na v 1.1 cDNA via transfection. Na v 1.1 mRNAs were quantified with qRT-PCR. By ANOVA and post hoc Sidak’s multiple comparisons, p = 0.003 for the comparison between control and shRNA1; p = 0.002 for the comparison between control and shRNA2. DOI: http://dx.doi.org/10.7554/eLife.13073.034

    Journal: eLife

    Article Title: A deleterious Nav1.1 mutation selectively impairs telencephalic inhibitory neurons derived from Dravet Syndrome patients

    doi: 10.7554/eLife.13073

    Figure Lengend Snippet: Transduction and RNAi efficiency of two human Nav1.1 lenti-shRNA clones. ( A ) Evaluating the transduction efficiency of the lentiviral particles were tested in HEK293T cells based on the expression of the tRFP reporter. Scale bars, 20 microns. ( B ) The RNAi efficiency of lenti-shRNA1 and lenti-shRNA2 relative to a non-silencing clone was determined by first transducing HEK293T cells with the lentiviral clones and next expressing the human Na v 1.1 cDNA via transfection. Na v 1.1 mRNAs were quantified with qRT-PCR. By ANOVA and post hoc Sidak’s multiple comparisons, p = 0.003 for the comparison between control and shRNA1; p = 0.002 for the comparison between control and shRNA2. DOI: http://dx.doi.org/10.7554/eLife.13073.034

    Article Snippet: Viral supernatants were concentrated using Lenti-X Concentrator (Clontech, Mountain View, California) and viral titers were determined with a Lenti-X qRT-PCR Titration Kit (Clontech).

    Techniques: Transduction, shRNA, Clone Assay, Expressing, Transfection, Quantitative RT-PCR