taqman gene expression assays Search Results


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  • 99
    Thermo Fisher taqman gene expression assays
    Real-time <t>PCR</t> showing the relative RNA expression level of Jam2 in mice with different genotypes. (A) <t>TaqMan</t> assay showed about 1.5-fold increase of Jam2 expression in Ts65Dn mice compared to WT. (B) TaqMan assay showed about 40% decrease of Jam2 expression
    Taqman Gene Expression Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52022 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gene expression master mix
    Effects of Target mRNA Structural and Positional Elements on Activities of <t>miRNA-mediated</t> Cleavage and Uridylation. ( a ) The plasmid construct of pLJ-T214 containing the EGFP reporter gene with two copies of predicted let-7a target sequences in its 3′UTR. ( b ) Cleaved mRNA 5′-fragments from let-7 targets T1 and T2 were detected by SLA–RT-PCR (upper gel panels) and SLA-qRT-PCR (lower panels). The predicted sizes of SLA–RT-PCR products derived from target sites T1 and T2 were 240-209 bp and 532-500 bp, respectively. Total RNAs were prepared from H1299 at 16, 40 and 72 h after pLJ-T214 transfection and RT reactions were performed using SLA-RT or 2U-SLA-RT primers. The corresponding target T2 cleavage and uridylation activities detected by SLA-qRT-PCR with T2 fragment specific PCR primers and <t>TaqMan</t> probe. ( c ) The plasmid construct of pLJ-T722. The 3′UTR of the EGFP reporter gene containing two copies of identical predicted let-7:target pairing sequences but with varied lengths and compositions of nt (underlined) at their 5′- and 3′-adjacent regions ( ST1 and ST2 ). ( d ) The effect of structural composition of target mRNA on let-7 miRNA–mediated mRNA cleavage activity as detected by SLA–RT-PCR assay (upper gel panels) and SLA-qRT-PCR (lower panels). Total RNAs were prepared from H1299 cells transfected with pLJ-T722 at 24 h, and SLA–RT-PCR was performed using SLA-RT primers. D50, a PCR primer specific to the pLJ-T722 transcript, was designed to bond at every 50 bases along the target sequences toward its 5′ end. The base numbers correspond to those shown in ( b ), and escaped bases are indicated by “•” between them. M: 0.03 μg of 1 kb DNA ladder. A Taqman probe-based qPCR assay was used to detect cleavage activity specific to target ST2 and a SYBR Green-based qPCR to detect both target ST1 and ST2 cleavage activities, respectively. All qPCR results were presented as relative fragment abundance (RFA). Each RFA value was represented as the mean of three independent experiments and error bars as standard errors to the mean. The target cleavage and uridylation activities in miRNA target regions were highlighted in red boxes.
    Taqman Gene Expression Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 21103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gene expression cells to ct kit
    Dose-dependent viral RNA amplification signal curve. Vero cells (2 × 10 4 /well) were cultured in a 96-well plate for 24 h in the absence of compounds. After incubation, the cells were infected with the indicated amount of STFSV and further incubated. After three days, the amount of intracellular viral RNA was determined by the real-time <t>RT-PCR</t> method using <t>TaqMan</t> Gene Expression Cells-to-CT™ Kit. The experiment was carried out in triplicate and means ± SD are shown.
    Taqman Gene Expression Cells To Ct Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqmangene expression assay
    M6P/IGF2R expression increases during monocyte differentiation to macrophages . ( A ) Cell‐surface expression of M6P/IGF2R on isolated PBMC and MACS‐enriched monocytes from healthy donors was evaluated with mAb MEM‐238‐AF647 and flow cytometry. In parallel, MOPC‐21‐AF647 was used as an isotype control mAb, displayed by the cut‐off gates. The same analysis was performed with macrophages differentiated from MACS‐sorted monocytes during a 7‐day culture with recombinant human M‐CSF (50 ng/ml) followed by 2 days resting in macrophage serum free medium. ( B ) Primary human monocytes and monocyte‐derived macrophages from (A) were lysed and RNA was extracted. cDNA was synthesized from total RNA and gene expression was measured by real‐time <t>PCR</t> as described in the Material and Methods section with <t>TaqMan</t> primer sets for human M6P/IGF2R and YWHAZ as endogenous control. The M6P/IGF2R mean expression values relative to that of monocytes ± SD from 3 donors is shown
    Taqmangene Expression Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1704 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher taqman gene expression mastermix
    M6P/IGF2R expression increases during monocyte differentiation to macrophages . ( A ) Cell‐surface expression of M6P/IGF2R on isolated PBMC and MACS‐enriched monocytes from healthy donors was evaluated with mAb MEM‐238‐AF647 and flow cytometry. In parallel, MOPC‐21‐AF647 was used as an isotype control mAb, displayed by the cut‐off gates. The same analysis was performed with macrophages differentiated from MACS‐sorted monocytes during a 7‐day culture with recombinant human M‐CSF (50 ng/ml) followed by 2 days resting in macrophage serum free medium. ( B ) Primary human monocytes and monocyte‐derived macrophages from (A) were lysed and RNA was extracted. cDNA was synthesized from total RNA and gene expression was measured by real‐time <t>PCR</t> as described in the Material and Methods section with <t>TaqMan</t> primer sets for human M6P/IGF2R and YWHAZ as endogenous control. The M6P/IGF2R mean expression values relative to that of monocytes ± SD from 3 donors is shown
    Taqman Gene Expression Mastermix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1288 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher custom taqman gene expression assays
    Levels of mRNA expression for the partner genes involved in chimeric transcripts in 26 paired samples of tumorous tissue (T) and non-cancerous renal cortex tissue (N) in the second cohort. mRNA expression was analyzed using custom <t>TaqMan</t> Gene Expression Assays on the 7500 Fast Real-Time PCR System (Life Technologies) employing the relative standard curve method. The probes and PCR primer sets used are summarized in Supporting Information Table S6. Experiments were performed in triplicate for each sample-primer set, and the mean value for the three experiments was used as the CT value. All CT values were normalized to that of GAPDH in the same sample. Levels of mRNA expression for the MMACHC, PTER, EPC2, ATXN7, FHIT, KIFAP3, CPEB1, MINPP1, TEX264, FAM107A , UPF3A, CDC16, MCCC1, CPSF3 , and ASAP2 genes were significantly reduced in T samples (shaded column) relative to N samples (white column). Bar, standard deviation.
    Custom Taqman Gene Expression Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 817 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Real-time PCR showing the relative RNA expression level of Jam2 in mice with different genotypes. (A) TaqMan assay showed about 1.5-fold increase of Jam2 expression in Ts65Dn mice compared to WT. (B) TaqMan assay showed about 40% decrease of Jam2 expression

    Journal: Genetics

    Article Title: Penetrance of Congenital Heart Disease in a Mouse Model of Down Syndrome Depends on a Trisomic Potentiator of a Disomic Modifier

    doi: 10.1534/genetics.116.188045

    Figure Lengend Snippet: Real-time PCR showing the relative RNA expression level of Jam2 in mice with different genotypes. (A) TaqMan assay showed about 1.5-fold increase of Jam2 expression in Ts65Dn mice compared to WT. (B) TaqMan assay showed about 40% decrease of Jam2 expression

    Article Snippet: PCR was carried out using Taqman Gene Expression Assays (Applied Biosystems, Foster City, CA).

    Techniques: Real-time Polymerase Chain Reaction, RNA Expression, Mouse Assay, TaqMan Assay, Expressing

    MAML1 promotes Runx2-mediated osteoblastic differentiation. A, C3H10T1/2 cells were transiently transfected with a Runx2 and/or MAML1 expression plasmids and cultured for 2 days. B, Total RNA was isolated and reverse-transcribed and TaqMan real-time PCR was performed to investigate the expression level of alkaline phosphatase gene, a marker of osteoblast.

    Journal: PLoS Genetics

    Article Title: MAML1 Enhances the Transcriptional Activity of Runx2 and Plays a Role in Bone Development

    doi: 10.1371/journal.pgen.1003132

    Figure Lengend Snippet: MAML1 promotes Runx2-mediated osteoblastic differentiation. A, C3H10T1/2 cells were transiently transfected with a Runx2 and/or MAML1 expression plasmids and cultured for 2 days. B, Total RNA was isolated and reverse-transcribed and TaqMan real-time PCR was performed to investigate the expression level of alkaline phosphatase gene, a marker of osteoblast.

    Article Snippet: Real-time PCR was performed using a TaqMan Gene Expression Assay, TaqMan Universal PCR Mix and the 7900HT Fast Real-Time PCR System (Applied Biosystems).

    Techniques: Transfection, Expressing, Cell Culture, Isolation, Real-time Polymerase Chain Reaction, Marker

    Expression of UL20, gK, and gB transcripts in corneas and TG of ocularly infected mice. WT and GODZ −/− mice were ocularly infected with 2 × 10 5 ). Briefly, the DNA template was serially diluted 10-fold so that 5 μl contained from 10 3 to 10 11 copies of each plasmid and then subjected to TaqMan PCR with the same set of primers. By comparing the normalized threshold cycle of each sample to the threshold cycle of the standard, the copy number for each reaction was determined. GAPDH expression was used to normalize the relative expression of each transcript in corneas and TG of infected mice. Shown are the means ± SEM from 6 corneas or TG. (A) UL20 transcript in corneas. (B) gK transcript in corneas. (C) gB transcript in corneas. (D) UL20 transcript in TG. (E) gK transcript in TG. (F) gB transcript in TG.

    Journal: Journal of Virology

    Article Title: The Absence of DHHC3 Affects Primary and Latent Herpes Simplex Virus 1 Infection

    doi: 10.1128/JVI.01599-17

    Figure Lengend Snippet: Expression of UL20, gK, and gB transcripts in corneas and TG of ocularly infected mice. WT and GODZ −/− mice were ocularly infected with 2 × 10 5 ). Briefly, the DNA template was serially diluted 10-fold so that 5 μl contained from 10 3 to 10 11 copies of each plasmid and then subjected to TaqMan PCR with the same set of primers. By comparing the normalized threshold cycle of each sample to the threshold cycle of the standard, the copy number for each reaction was determined. GAPDH expression was used to normalize the relative expression of each transcript in corneas and TG of infected mice. Shown are the means ± SEM from 6 corneas or TG. (A) UL20 transcript in corneas. (B) gK transcript in corneas. (C) gB transcript in corneas. (D) UL20 transcript in TG. (E) gK transcript in TG. (F) gB transcript in TG.

    Article Snippet: The expression levels of gB, gK, UL20, and LAT, as well as the expression of the endogenous control GAPDH gene, were evaluated using a commercially available TaqMan gene expression assay kit (Applied Biosystems, Foster City, CA) containing optimized primer and probe concentrations.

    Techniques: Expressing, Infection, Mouse Assay, Plasmid Preparation, Polymerase Chain Reaction

    NFAT1 inhibits Cyclin E1 and E2 expression in primary B lymphocytes. (A-C) B lymphocytes were purified from naive NFAT1+/+ and NFAT1−/− mice and stimulated with PMA (10 nM) and Ionomycin (1 μM) for 24 hours (A and B) or 48 hours (C). (A) Total RNA was extracted and the expression of Myc family members (left panel) and Cyclin family members (middle and right panels) were analyzed by RNase Protection Assay. Transcript levels were analyzed by autoradiography, and RNA loading was estimated by measuring the intensities of 2 housekeeping genes (L32 and GAPDH). (B) Total RNA was extracted and Cyclin E1 and Cyclin E2 expression levels were analyzed by real-time PCR using TaqMan Gene Expression assays. HPRT expression was used for normalization. (C) Total protein lysates were obtained and Cyclin E1, Cyclin E2, and GAPDH protein levels were detected by western blot. Cyclin E protein levels were normalized to GAPDH and are indicated below blots. All results were obtained from a pool of 3 mice, and are representative of at least 2 independent experiments. Results are expressed as mean and error bars represent SEM. Asterisks indicate significance levels compared to controls (* p

    Journal: Cell Cycle

    Article Title: NFAT1 transcription factor regulates cell cycle progression and cyclin E expression in B lymphocytes

    doi: 10.1080/15384101.2016.1203485

    Figure Lengend Snippet: NFAT1 inhibits Cyclin E1 and E2 expression in primary B lymphocytes. (A-C) B lymphocytes were purified from naive NFAT1+/+ and NFAT1−/− mice and stimulated with PMA (10 nM) and Ionomycin (1 μM) for 24 hours (A and B) or 48 hours (C). (A) Total RNA was extracted and the expression of Myc family members (left panel) and Cyclin family members (middle and right panels) were analyzed by RNase Protection Assay. Transcript levels were analyzed by autoradiography, and RNA loading was estimated by measuring the intensities of 2 housekeeping genes (L32 and GAPDH). (B) Total RNA was extracted and Cyclin E1 and Cyclin E2 expression levels were analyzed by real-time PCR using TaqMan Gene Expression assays. HPRT expression was used for normalization. (C) Total protein lysates were obtained and Cyclin E1, Cyclin E2, and GAPDH protein levels were detected by western blot. Cyclin E protein levels were normalized to GAPDH and are indicated below blots. All results were obtained from a pool of 3 mice, and are representative of at least 2 independent experiments. Results are expressed as mean and error bars represent SEM. Asterisks indicate significance levels compared to controls (* p

    Article Snippet: Relative quantification of CCNE1 and CCNE2 transcripts was performed by real-time PCR using a 7500 Real Time PCR System (Applied Biosystems) and the following TaqMan Gene Expression Assay kits: CCNE1, Mm00432367_m1; CCNE2, Mm00438077_m1; and HPRT, Mm00446968_m1 (Thermo Fisher Scientific).

    Techniques: Expressing, Purification, Mouse Assay, Rnase Protection Assay, Autoradiography, Real-time Polymerase Chain Reaction, Western Blot

    TaqMan® PCR validation of microarray experiments. Profile charts of gene expression levels comparing results obtained by microarray analysis ( n = 25) to TaqMan® PCR analysis ( n = 20) for six genes. Plots for those genes with multiple probes are also displayed where appropriate

    Journal: Virchows Archiv

    Article Title: Expression microarray analysis of papillary thyroid carcinoma and benign thyroid tissue: emphasis on the follicular variant and potential markers of malignancy

    doi: 10.1007/s00428-006-0348-5

    Figure Lengend Snippet: TaqMan® PCR validation of microarray experiments. Profile charts of gene expression levels comparing results obtained by microarray analysis ( n = 25) to TaqMan® PCR analysis ( n = 20) for six genes. Plots for those genes with multiple probes are also displayed where appropriate

    Article Snippet: Primers and probes for TaqMan® PCR were obtained by using Applied Biosystems’ pre-designed TaqMan® Gene Expression Assays.

    Techniques: Polymerase Chain Reaction, Microarray, Expressing

    Hypoxia-induced modification of PLK promoter methylation in HCC cells. (a) Promoter methylation status of the plks examined in HCC-derived cells HepG2 and Hep3B; U = unmethylated, M = methylated. Fully methylated HeLa DNA was used as a positive control (+M), no template was added to the negative control (−M). (b) Post hypoxia, PLK4 transcripts were assessed via qPCR in RNA extracted from HCC cells. All qPCR data is representative of the mean value of three independent experiments and error bars represent +/− SD. (c) PLK protein levels were examined post treatment from whole cell lysates. Actin was used as a loading control. (−) represents lysates from untreated cells, (+) lysates from cells grown in the presence of hypoxia. (d) Quantification of protein levels using densitometry. Levels have been normalized to the respective untreated controls. Data is representative of the mean value of three independent experiments and error bars represent +/− SD. (e) The fold change of PLK1 transcripts as determined by qPCR. Values normalized to the respective untreated sample. (f) PLK2 and PLK3 analyzed and fold changed determine by normalization to the respective untreated samples. (g) Hif1α transcripts post hypoxia were determine by real-time PCR using a Taqman probe.

    Journal: PLoS ONE

    Article Title: p53-Dependent and Cell Specific Epigenetic Regulation of the Polo-like kinases under Oxidative Stress

    doi: 10.1371/journal.pone.0087918

    Figure Lengend Snippet: Hypoxia-induced modification of PLK promoter methylation in HCC cells. (a) Promoter methylation status of the plks examined in HCC-derived cells HepG2 and Hep3B; U = unmethylated, M = methylated. Fully methylated HeLa DNA was used as a positive control (+M), no template was added to the negative control (−M). (b) Post hypoxia, PLK4 transcripts were assessed via qPCR in RNA extracted from HCC cells. All qPCR data is representative of the mean value of three independent experiments and error bars represent +/− SD. (c) PLK protein levels were examined post treatment from whole cell lysates. Actin was used as a loading control. (−) represents lysates from untreated cells, (+) lysates from cells grown in the presence of hypoxia. (d) Quantification of protein levels using densitometry. Levels have been normalized to the respective untreated controls. Data is representative of the mean value of three independent experiments and error bars represent +/− SD. (e) The fold change of PLK1 transcripts as determined by qPCR. Values normalized to the respective untreated sample. (f) PLK2 and PLK3 analyzed and fold changed determine by normalization to the respective untreated samples. (g) Hif1α transcripts post hypoxia were determine by real-time PCR using a Taqman probe.

    Article Snippet: Real time PCRs were carried out on an ABI 7300 machine using Taqman gene expression probes for mouse Plk1 , Plk4 , and HIF1α ; and human PLK1-PLK4 , and HIF1α (Applied biosystems).

    Techniques: Modification, Methylation, Derivative Assay, Positive Control, Negative Control, Real-time Polymerase Chain Reaction

    qPCR verification of differentially expressed phototransduction genes observed in RNA-seq data at 72 hpf. SuperScript III was used to generate cDNA from total RNA. Message level was determined on a 7900HT Sequence Detection System using predesigned TaqMan

    Journal: Drug Metabolism and Disposition

    Article Title: Inhibition of SULT4A1 Expression Induces Up-Regulation of Phototransduction Gene Expression in 72-Hour Postfertilization Zebrafish Larvae

    doi: 10.1124/dmd.114.057042

    Figure Lengend Snippet: qPCR verification of differentially expressed phototransduction genes observed in RNA-seq data at 72 hpf. SuperScript III was used to generate cDNA from total RNA. Message level was determined on a 7900HT Sequence Detection System using predesigned TaqMan

    Article Snippet: RNA concentration was determined using a NanoDrop ND-1000 spectrophotometer, and SuperScript III (Invitrogen, Carlsbad, California) was used to generate cDNA using 200-ng RNA from each sample. qPCR experiments comparing the following genes were carried out using predesigned TaqMan gene expression assays from Life Technologies with an Applied Biosystems 7900HT Sequence Detection System (Applied Biosystems, Foster City, CA)—SULT4A1, assay ID: Dr03078008_g1; retinal pigment epithelium–derived rhodopsin homolog (rrh), assay ID: Dr03108770_m1; G protein–coupled receptor kinase 1b (grk1b), assay ID: Dr03128502_m1; guanylate cyclase activator 1e (guca1e), assay ID: Dr03094375_m1; and opsin 1, medium-wave-sensitive, 1 (opn1mw1), assay ID: Dr03079939_g1.

    Techniques: Real-time Polymerase Chain Reaction, RNA Sequencing Assay, Sequencing

    qPCR analysis of dorsal root ganglions (DRGs) from scLRP1 −/− and scLRP1 +/+ mice show upregulation of regeneration associated genes (RAGs) before and after injury Taqman qPCR analysis of ( A ) uninjured DRGs showing CREB3, ATF3, STAT3 and SPRR1A in scLRP1 −/− mice (n=3–5 mice per genotype; *p

    Journal: Glia

    Article Title: Schwann Cells Regulate Sensory Neuron Gene Expression Before and After Peripheral Nerve Injury

    doi: 10.1002/glia.23325

    Figure Lengend Snippet: qPCR analysis of dorsal root ganglions (DRGs) from scLRP1 −/− and scLRP1 +/+ mice show upregulation of regeneration associated genes (RAGs) before and after injury Taqman qPCR analysis of ( A ) uninjured DRGs showing CREB3, ATF3, STAT3 and SPRR1A in scLRP1 −/− mice (n=3–5 mice per genotype; *p

    Article Snippet: RNA was reverse-transcribed using the iScript cDNA synthesis kit (Bio-Rad). qPCR was performed using TaqMan® gene expression products and an AB Step one Plus Real-Time PCR System (Applied Biosystems), as previously described ( ; ).

    Techniques: Real-time Polymerase Chain Reaction, Mouse Assay

    Activation of FOXO pathway and TXNIP induction. A : HepG2 and HuH-7 cells were treated with 250 μM MK-801 or NBQX for 12 h. cDNA was synthesized and real-time quantitative PCR was carried out using Taqman gene expression assay primers. Each reaction was performed in duplicate. The β-actin gene was used to normalize across assays and runs, and the threshold value (Ct) for each sample was used to determine gene expression levels. Each bar represents the mean ± SD of at least three replicates. B : HepG2 cells were treated with 250 μM MK-801 and expression of TXNIP and p27 was measured by real-time quantitative PCR for 72 h. Each bar represents the mean ± SD of at least three replicates. C : HepG2 cells were treated with 250 μM MK-801 for 0 to 96 h and Western blot analysis was performed using indicated antibodies. D : HepG2 cells were treated with 250 μM MK-801 for 0 to 60 min and Western blot analysis was carried out using indicated antibodies. The molecular weight of dephosphorylated band corresponded to Thr24 of FOXO1 E : FOXO1-pAcGFP-N1 plasmid was transfected to HepG2 cells and treated with or without 250 μM of MK-801. Nuclear translocation of FOXO1-GFP protein was observed with Olympus LX71 microscope.

    Journal: BMC Cancer

    Article Title: FOXO/TXNIP pathway is involved in the suppression of hepatocellular carcinoma growth by glutamate antagonist MK-801

    doi: 10.1186/1471-2407-13-468

    Figure Lengend Snippet: Activation of FOXO pathway and TXNIP induction. A : HepG2 and HuH-7 cells were treated with 250 μM MK-801 or NBQX for 12 h. cDNA was synthesized and real-time quantitative PCR was carried out using Taqman gene expression assay primers. Each reaction was performed in duplicate. The β-actin gene was used to normalize across assays and runs, and the threshold value (Ct) for each sample was used to determine gene expression levels. Each bar represents the mean ± SD of at least three replicates. B : HepG2 cells were treated with 250 μM MK-801 and expression of TXNIP and p27 was measured by real-time quantitative PCR for 72 h. Each bar represents the mean ± SD of at least three replicates. C : HepG2 cells were treated with 250 μM MK-801 for 0 to 96 h and Western blot analysis was performed using indicated antibodies. D : HepG2 cells were treated with 250 μM MK-801 for 0 to 60 min and Western blot analysis was carried out using indicated antibodies. The molecular weight of dephosphorylated band corresponded to Thr24 of FOXO1 E : FOXO1-pAcGFP-N1 plasmid was transfected to HepG2 cells and treated with or without 250 μM of MK-801. Nuclear translocation of FOXO1-GFP protein was observed with Olympus LX71 microscope.

    Article Snippet: Real-time quantitative PCR was carried out using Taqman gene expression assay primers and a 7300 real-time PCR system (Applied Biosystems).

    Techniques: Activation Assay, Synthesized, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Molecular Weight, Plasmid Preparation, Transfection, Translocation Assay, Microscopy

    Analysis of the alternatively activated (M2) macrophage markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw (arthritis score ≥ 3) on D7. Relative mRNA expression of CD163 (A) and CD206 (B) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each datum point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Journal: Prostaglandins, leukotrienes, and essential fatty acids

    Article Title: ANTI-INFLAMMATORY PROPERTIES OF PROSTAGLANDIN E2: DELETION OF MICROSOMAL PROSTAGLANDIN E SYNTHASE-1 EXACERBATES NON-IMMUNE INFLAMMATORY ARTHRITIS IN MICE

    doi: 10.1016/j.plefa.2013.08.003

    Figure Lengend Snippet: Analysis of the alternatively activated (M2) macrophage markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw (arthritis score ≥ 3) on D7. Relative mRNA expression of CD163 (A) and CD206 (B) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each datum point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Article Snippet: Particular genes were detected using the respective Taq Man Gene Expression Assays (Applied Biosystems) on a 7300 Real-Time RT-PCR system from the same manufacturer by relative quantitation employing glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the reference gene.

    Techniques: Quantitative RT-PCR, Isolation, Expressing

    Analysis of inflammatory markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw on D7. Relative mRNA expression of MPO (A), IL-6 (B), and PTGS2 (C) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each datum point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Journal: Prostaglandins, leukotrienes, and essential fatty acids

    Article Title: ANTI-INFLAMMATORY PROPERTIES OF PROSTAGLANDIN E2: DELETION OF MICROSOMAL PROSTAGLANDIN E SYNTHASE-1 EXACERBATES NON-IMMUNE INFLAMMATORY ARTHRITIS IN MICE

    doi: 10.1016/j.plefa.2013.08.003

    Figure Lengend Snippet: Analysis of inflammatory markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw on D7. Relative mRNA expression of MPO (A), IL-6 (B), and PTGS2 (C) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each datum point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Article Snippet: Particular genes were detected using the respective Taq Man Gene Expression Assays (Applied Biosystems) on a 7300 Real-Time RT-PCR system from the same manufacturer by relative quantitation employing glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the reference gene.

    Techniques: Quantitative RT-PCR, Isolation, Expressing

    Analysis of the classically activated (M1) macrophage markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw (arthritis score ≥ 3) on D7. Relative mRNA expression of CCL3 (A) and NOS2 (B) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each data point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Journal: Prostaglandins, leukotrienes, and essential fatty acids

    Article Title: ANTI-INFLAMMATORY PROPERTIES OF PROSTAGLANDIN E2: DELETION OF MICROSOMAL PROSTAGLANDIN E SYNTHASE-1 EXACERBATES NON-IMMUNE INFLAMMATORY ARTHRITIS IN MICE

    doi: 10.1016/j.plefa.2013.08.003

    Figure Lengend Snippet: Analysis of the classically activated (M1) macrophage markers in arthritic paws of WT and mPGES-1 KO animals by real-time RT-PCR. Total RNA was obtained from a synovial capsule isolated from the front arthritic paw (arthritis score ≥ 3) on D7. Relative mRNA expression of CCL3 (A) and NOS2 (B) was measured using the respective Taq Man Gene Expression Assays from Applied Biosystems. Each data point represents a single animal. Two animal groups were compared by Student t -test with the P ≤ 0.05 considered to be significant (n=9).

    Article Snippet: Particular genes were detected using the respective Taq Man Gene Expression Assays (Applied Biosystems) on a 7300 Real-Time RT-PCR system from the same manufacturer by relative quantitation employing glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the reference gene.

    Techniques: Quantitative RT-PCR, Isolation, Expressing

    Altered miRNAs in neonatal vs. adult mouse lung. A : Venn diagram of altered miRNAs in neonatal vs. adult lungs. Fifty miRNAs increased more than twofold, and 17 miRNAs decreased by more than half in neonates exposed to hyperoxia. In adults exposed to hyperoxia, 21 miRNAs increased more than twofold, and 12 miRNAs decreased by more than half. B : this Venn diagram of miRNA related to Bach1 shows no altered miRNAs common to either neonatal or adult lungs exposed to hyperoxia on the basis of array data. C : selected miRNAs from the miRNA expression profile were validated by RT-quantitative PCR (qPCR). D : correlation between data from microarray and RT-PCR for selected miRNAs was performed using lineal regression. The x -axis represents fold in log 2 by qPCR; the y -axis represents fold in log 2 by TaqMan low-density array data.

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: MiR-196a regulates heme oxygenase-1 by silencing Bach1 in the neonatal mouse lung

    doi: 10.1152/ajplung.00428.2015

    Figure Lengend Snippet: Altered miRNAs in neonatal vs. adult mouse lung. A : Venn diagram of altered miRNAs in neonatal vs. adult lungs. Fifty miRNAs increased more than twofold, and 17 miRNAs decreased by more than half in neonates exposed to hyperoxia. In adults exposed to hyperoxia, 21 miRNAs increased more than twofold, and 12 miRNAs decreased by more than half. B : this Venn diagram of miRNA related to Bach1 shows no altered miRNAs common to either neonatal or adult lungs exposed to hyperoxia on the basis of array data. C : selected miRNAs from the miRNA expression profile were validated by RT-quantitative PCR (qPCR). D : correlation between data from microarray and RT-PCR for selected miRNAs was performed using lineal regression. The x -axis represents fold in log 2 by qPCR; the y -axis represents fold in log 2 by TaqMan low-density array data.

    Article Snippet: Steady-state mRNA and miRNA levels were evaluated by quantitative real-time PCR using a TaqMan gene expression system (Applied Biosystems).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Microarray, Reverse Transcription Polymerase Chain Reaction, TLDA Assay

    Quantitative RT-PCR analysis of TNF-α regulated genes in U937 cells after sequential treatment with PGSE and TNF-α . U937 cells (1 × 10 6 ) were pretreated with or without 3 mg/ml PGSE for 24 hours and followed by 20 units/ml of TNF-α for 2 hours. DNase-treated RNA samples were reverse transcribed and the levels of mRNA induction of (A) CXCL-10, (B) IL-8 and (C) TNFAIP3 as well as the reference gene 18S rRNA were determined by gene-specific TaqMan assays as described in Methods. The levels of induction were relative to the untreated cells. Values represent the average ± SD of three independent experiments and statistically analyzed by two tailed, paired t-test. *: p

    Journal: Journal of Translational Medicine

    Article Title: Bioactivity-guided identification and cell signaling technology to delineate the immunomodulatory effects of Panax ginseng on human promonocytic U937 cells

    doi: 10.1186/1479-5876-7-34

    Figure Lengend Snippet: Quantitative RT-PCR analysis of TNF-α regulated genes in U937 cells after sequential treatment with PGSE and TNF-α . U937 cells (1 × 10 6 ) were pretreated with or without 3 mg/ml PGSE for 24 hours and followed by 20 units/ml of TNF-α for 2 hours. DNase-treated RNA samples were reverse transcribed and the levels of mRNA induction of (A) CXCL-10, (B) IL-8 and (C) TNFAIP3 as well as the reference gene 18S rRNA were determined by gene-specific TaqMan assays as described in Methods. The levels of induction were relative to the untreated cells. Values represent the average ± SD of three independent experiments and statistically analyzed by two tailed, paired t-test. *: p

    Article Snippet: Briefly, DNase-treated RNA samples were reverse transcribed using TaqMan reverse transcription reagent kit (Applied Biosystems) and the levels of CXCL-10, IL-8 and TNFAIP3 mRNA as well as the reference gene 18S rRNA were assayed by the gene-specific TaqMan gene expression assays (Applied Biosystems).

    Techniques: Quantitative RT-PCR, Two Tailed Test

    Effects of Target mRNA Structural and Positional Elements on Activities of miRNA-mediated Cleavage and Uridylation. ( a ) The plasmid construct of pLJ-T214 containing the EGFP reporter gene with two copies of predicted let-7a target sequences in its 3′UTR. ( b ) Cleaved mRNA 5′-fragments from let-7 targets T1 and T2 were detected by SLA–RT-PCR (upper gel panels) and SLA-qRT-PCR (lower panels). The predicted sizes of SLA–RT-PCR products derived from target sites T1 and T2 were 240-209 bp and 532-500 bp, respectively. Total RNAs were prepared from H1299 at 16, 40 and 72 h after pLJ-T214 transfection and RT reactions were performed using SLA-RT or 2U-SLA-RT primers. The corresponding target T2 cleavage and uridylation activities detected by SLA-qRT-PCR with T2 fragment specific PCR primers and TaqMan probe. ( c ) The plasmid construct of pLJ-T722. The 3′UTR of the EGFP reporter gene containing two copies of identical predicted let-7:target pairing sequences but with varied lengths and compositions of nt (underlined) at their 5′- and 3′-adjacent regions ( ST1 and ST2 ). ( d ) The effect of structural composition of target mRNA on let-7 miRNA–mediated mRNA cleavage activity as detected by SLA–RT-PCR assay (upper gel panels) and SLA-qRT-PCR (lower panels). Total RNAs were prepared from H1299 cells transfected with pLJ-T722 at 24 h, and SLA–RT-PCR was performed using SLA-RT primers. D50, a PCR primer specific to the pLJ-T722 transcript, was designed to bond at every 50 bases along the target sequences toward its 5′ end. The base numbers correspond to those shown in ( b ), and escaped bases are indicated by “•” between them. M: 0.03 μg of 1 kb DNA ladder. A Taqman probe-based qPCR assay was used to detect cleavage activity specific to target ST2 and a SYBR Green-based qPCR to detect both target ST1 and ST2 cleavage activities, respectively. All qPCR results were presented as relative fragment abundance (RFA). Each RFA value was represented as the mean of three independent experiments and error bars as standard errors to the mean. The target cleavage and uridylation activities in miRNA target regions were highlighted in red boxes.

    Journal: Scientific Reports

    Article Title: MicroRNA-mediated target mRNA cleavage and 3′-uridylation in human cells

    doi: 10.1038/srep30242

    Figure Lengend Snippet: Effects of Target mRNA Structural and Positional Elements on Activities of miRNA-mediated Cleavage and Uridylation. ( a ) The plasmid construct of pLJ-T214 containing the EGFP reporter gene with two copies of predicted let-7a target sequences in its 3′UTR. ( b ) Cleaved mRNA 5′-fragments from let-7 targets T1 and T2 were detected by SLA–RT-PCR (upper gel panels) and SLA-qRT-PCR (lower panels). The predicted sizes of SLA–RT-PCR products derived from target sites T1 and T2 were 240-209 bp and 532-500 bp, respectively. Total RNAs were prepared from H1299 at 16, 40 and 72 h after pLJ-T214 transfection and RT reactions were performed using SLA-RT or 2U-SLA-RT primers. The corresponding target T2 cleavage and uridylation activities detected by SLA-qRT-PCR with T2 fragment specific PCR primers and TaqMan probe. ( c ) The plasmid construct of pLJ-T722. The 3′UTR of the EGFP reporter gene containing two copies of identical predicted let-7:target pairing sequences but with varied lengths and compositions of nt (underlined) at their 5′- and 3′-adjacent regions ( ST1 and ST2 ). ( d ) The effect of structural composition of target mRNA on let-7 miRNA–mediated mRNA cleavage activity as detected by SLA–RT-PCR assay (upper gel panels) and SLA-qRT-PCR (lower panels). Total RNAs were prepared from H1299 cells transfected with pLJ-T722 at 24 h, and SLA–RT-PCR was performed using SLA-RT primers. D50, a PCR primer specific to the pLJ-T722 transcript, was designed to bond at every 50 bases along the target sequences toward its 5′ end. The base numbers correspond to those shown in ( b ), and escaped bases are indicated by “•” between them. M: 0.03 μg of 1 kb DNA ladder. A Taqman probe-based qPCR assay was used to detect cleavage activity specific to target ST2 and a SYBR Green-based qPCR to detect both target ST1 and ST2 cleavage activities, respectively. All qPCR results were presented as relative fragment abundance (RFA). Each RFA value was represented as the mean of three independent experiments and error bars as standard errors to the mean. The target cleavage and uridylation activities in miRNA target regions were highlighted in red boxes.

    Article Snippet: High-capacity cDNA first-strand synthesis kit, TaqMan Gene Expression Master Mix and TaqMan probes for miRNA quantification were purchased from Applied Biosystems (Life Technologies).

    Techniques: Plasmid Preparation, Construct, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Derivative Assay, Transfection, Polymerase Chain Reaction, Activity Assay, Real-time Polymerase Chain Reaction, SYBR Green Assay

    Reduction of a viral-host fusion transcript 4 in SiHa cells promotes cell proliferation. A . Major viral-host fusion transcripts in SiHa cells. Arrows are the primers used for RT-PCR detection of the fusion transcripts. A synthetic siRNA specifically targeting the transcript 4 (si-Junction) was custom-designed. B - D . Reduction of the transcript 4 expression in SiHa cells led to reduction of p53 protein. SiHa cells were harvested for total cell protein and RNA preparation 48 h after siRNA transfection. Knockdown efficiency of the transcript 4 was evaluated by RT-PCR using a primer set of F1+F12 (B). Expression of the transcript 2 and GAPDH served as a sample loading control. Knockdown efficiency of the transcript 4 was also evaluated by RT-qPCR using a virus-host specific junction TaqMan probe (C). Effect of transcript 4 knockdown on p53 expression was examined by Western blot with □-tubulin serving as a sample loading control (D). E. Enhancement of SiHa cell proliferation by knockdown of transcript 4 expression. SiHa cells at 24 h after plating were transfected with 40 nM of siRNA (arrow, si-Junction or si-Control) and the cell proliferation was examined at the indicated time (h) after siRNA transfection by averaged cell number mean ± SE from three experimental repeats. **, p

    Journal: bioRxiv

    Article Title: Expression of HPV oncogenes from a single integration site in cervical cancer cell lines

    doi: 10.1101/2020.06.25.171959

    Figure Lengend Snippet: Reduction of a viral-host fusion transcript 4 in SiHa cells promotes cell proliferation. A . Major viral-host fusion transcripts in SiHa cells. Arrows are the primers used for RT-PCR detection of the fusion transcripts. A synthetic siRNA specifically targeting the transcript 4 (si-Junction) was custom-designed. B - D . Reduction of the transcript 4 expression in SiHa cells led to reduction of p53 protein. SiHa cells were harvested for total cell protein and RNA preparation 48 h after siRNA transfection. Knockdown efficiency of the transcript 4 was evaluated by RT-PCR using a primer set of F1+F12 (B). Expression of the transcript 2 and GAPDH served as a sample loading control. Knockdown efficiency of the transcript 4 was also evaluated by RT-qPCR using a virus-host specific junction TaqMan probe (C). Effect of transcript 4 knockdown on p53 expression was examined by Western blot with □-tubulin serving as a sample loading control (D). E. Enhancement of SiHa cell proliferation by knockdown of transcript 4 expression. SiHa cells at 24 h after plating were transfected with 40 nM of siRNA (arrow, si-Junction or si-Control) and the cell proliferation was examined at the indicated time (h) after siRNA transfection by averaged cell number mean ± SE from three experimental repeats. **, p

    Article Snippet: RT-qPCR was carried out with total RNA isolated from SiHa cells using a TaqMan Gene Expression kit (Thermo Fisher Scientific, #4369016).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Quantitative RT-PCR, Western Blot

    Dose-dependent viral RNA amplification signal curve. Vero cells (2 × 10 4 /well) were cultured in a 96-well plate for 24 h in the absence of compounds. After incubation, the cells were infected with the indicated amount of STFSV and further incubated. After three days, the amount of intracellular viral RNA was determined by the real-time RT-PCR method using TaqMan Gene Expression Cells-to-CT™ Kit. The experiment was carried out in triplicate and means ± SD are shown.

    Journal: Antiviral Chemistry & Chemotherapy

    Article Title: Establishment of an antiviral assay system and identification of severe fever with thrombocytopenia syndrome virus inhibitors

    doi: 10.1177/2040206617740303

    Figure Lengend Snippet: Dose-dependent viral RNA amplification signal curve. Vero cells (2 × 10 4 /well) were cultured in a 96-well plate for 24 h in the absence of compounds. After incubation, the cells were infected with the indicated amount of STFSV and further incubated. After three days, the amount of intracellular viral RNA was determined by the real-time RT-PCR method using TaqMan Gene Expression Cells-to-CT™ Kit. The experiment was carried out in triplicate and means ± SD are shown.

    Article Snippet: The infected cells exposed to various concentrations of test compounds can be treated with lysis buffer after washing with PBS and subjected to real-time RT-PCR without extracting RNA, when TaqMan Gene Expression Cells-to-CT™ Kit is used for the assay.

    Techniques: Amplification, Cell Culture, Incubation, Infection, Quantitative RT-PCR, Expressing

    M6P/IGF2R expression increases during monocyte differentiation to macrophages . ( A ) Cell‐surface expression of M6P/IGF2R on isolated PBMC and MACS‐enriched monocytes from healthy donors was evaluated with mAb MEM‐238‐AF647 and flow cytometry. In parallel, MOPC‐21‐AF647 was used as an isotype control mAb, displayed by the cut‐off gates. The same analysis was performed with macrophages differentiated from MACS‐sorted monocytes during a 7‐day culture with recombinant human M‐CSF (50 ng/ml) followed by 2 days resting in macrophage serum free medium. ( B ) Primary human monocytes and monocyte‐derived macrophages from (A) were lysed and RNA was extracted. cDNA was synthesized from total RNA and gene expression was measured by real‐time PCR as described in the Material and Methods section with TaqMan primer sets for human M6P/IGF2R and YWHAZ as endogenous control. The M6P/IGF2R mean expression values relative to that of monocytes ± SD from 3 donors is shown

    Journal: Journal of Leukocyte Biology

    Article Title: The mannose 6‐phosphate/insulin‐like growth factor 2 receptor mediates plasminogen‐induced efferocytosis, et al. The mannose 6‐phosphate/insulin‐like growth factor 2 receptor mediates plasminogen‐induced efferocytosis

    doi: 10.1002/JLB.1AB0417-160RR

    Figure Lengend Snippet: M6P/IGF2R expression increases during monocyte differentiation to macrophages . ( A ) Cell‐surface expression of M6P/IGF2R on isolated PBMC and MACS‐enriched monocytes from healthy donors was evaluated with mAb MEM‐238‐AF647 and flow cytometry. In parallel, MOPC‐21‐AF647 was used as an isotype control mAb, displayed by the cut‐off gates. The same analysis was performed with macrophages differentiated from MACS‐sorted monocytes during a 7‐day culture with recombinant human M‐CSF (50 ng/ml) followed by 2 days resting in macrophage serum free medium. ( B ) Primary human monocytes and monocyte‐derived macrophages from (A) were lysed and RNA was extracted. cDNA was synthesized from total RNA and gene expression was measured by real‐time PCR as described in the Material and Methods section with TaqMan primer sets for human M6P/IGF2R and YWHAZ as endogenous control. The M6P/IGF2R mean expression values relative to that of monocytes ± SD from 3 donors is shown

    Article Snippet: Quantitative PCR was carried out in duplicates using the TaqMan® Gene Expression Assay System (Invitrogen) in a CFX96 Touch Real Time PCR Detection System (Bio‐Rad, Hercules, CA).

    Techniques: Expressing, Isolation, Magnetic Cell Separation, Flow Cytometry, Cytometry, Recombinant, Derivative Assay, Synthesized, Real-time Polymerase Chain Reaction

    Altered expression of TRP genes by paclitaxel. A . Screening of expression of TRP genes by DNA microarray. B . TRPV4 validaton by RT-PCR. TRPV4 gene, whose change in expression was significant in DNA microarray, was determined by TaqMan Gene Expression Assay. † P

    Journal: Molecular Pain

    Article Title: The prophylactic effects of a traditional Japanese medicine, goshajinkigan, on paclitaxel-induced peripheral neuropathy and its mechanism of action

    doi: 10.1186/1744-8069-10-61

    Figure Lengend Snippet: Altered expression of TRP genes by paclitaxel. A . Screening of expression of TRP genes by DNA microarray. B . TRPV4 validaton by RT-PCR. TRPV4 gene, whose change in expression was significant in DNA microarray, was determined by TaqMan Gene Expression Assay. † P

    Article Snippet: Before qPCR, reverse transcription of 500 ng of total RNA was performed using SuperscriptR VILO® cDNA synthesis kit (Life Technologies), qPCR reactions were set-up in 20 μl volumes with 50-fold cDNA dilutions, 20× TaqMan® gene expression assay mix, 2× TaqMan® Universal PCR master mix II (Life Technologies), and dH2 O. PCRs were performed in quadruplicate following the manufacture protocols on an model ABI Prism 7000 real-time PCR system (Life Technologies) using the following protocol: an initial denaturation and polymerase activation step for 10 min at 95°C, followed by 40 cycles of denaturation at 95°C for 15 s and 60°C for 1 min. Actb was used as a reference gene to normalize between samples.

    Techniques: Expressing, Microarray, Reverse Transcription Polymerase Chain Reaction

    Levels of mRNA expression for the partner genes involved in chimeric transcripts in 26 paired samples of tumorous tissue (T) and non-cancerous renal cortex tissue (N) in the second cohort. mRNA expression was analyzed using custom TaqMan Gene Expression Assays on the 7500 Fast Real-Time PCR System (Life Technologies) employing the relative standard curve method. The probes and PCR primer sets used are summarized in Supporting Information Table S6. Experiments were performed in triplicate for each sample-primer set, and the mean value for the three experiments was used as the CT value. All CT values were normalized to that of GAPDH in the same sample. Levels of mRNA expression for the MMACHC, PTER, EPC2, ATXN7, FHIT, KIFAP3, CPEB1, MINPP1, TEX264, FAM107A , UPF3A, CDC16, MCCC1, CPSF3 , and ASAP2 genes were significantly reduced in T samples (shaded column) relative to N samples (white column). Bar, standard deviation.

    Journal: Genes, Chromosomes & Cancer

    Article Title: Comprehensive Exploration of Novel Chimeric Transcripts in Clear Cell Renal Cell Carcinomas Using Whole Transcriptome Analysis

    doi: 10.1002/gcc.22211

    Figure Lengend Snippet: Levels of mRNA expression for the partner genes involved in chimeric transcripts in 26 paired samples of tumorous tissue (T) and non-cancerous renal cortex tissue (N) in the second cohort. mRNA expression was analyzed using custom TaqMan Gene Expression Assays on the 7500 Fast Real-Time PCR System (Life Technologies) employing the relative standard curve method. The probes and PCR primer sets used are summarized in Supporting Information Table S6. Experiments were performed in triplicate for each sample-primer set, and the mean value for the three experiments was used as the CT value. All CT values were normalized to that of GAPDH in the same sample. Levels of mRNA expression for the MMACHC, PTER, EPC2, ATXN7, FHIT, KIFAP3, CPEB1, MINPP1, TEX264, FAM107A , UPF3A, CDC16, MCCC1, CPSF3 , and ASAP2 genes were significantly reduced in T samples (shaded column) relative to N samples (white column). Bar, standard deviation.

    Article Snippet: Quantitative RT-PCR Analysis cDNA was reverse transcribed from total RNA (500 ng) of the 26 paired T and N samples of the second cohort using random primers and Superscript III RNase H− Reverse Transcriptase (Life Technologies). mRNA expression was analyzed using custom TaqMan Gene Expression Assays and TaqMan Fast Advanced Master Mix (Life Technologies) on a 7500 Fast Real-Time PCR System (Life Technologies) employing the relative standard curve method.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Standard Deviation

    Gene expression of M-CSFR in 4 rainbow trout immune tissues (AK, PBL, SPL, and PK) from YC13 and/or YC15, using RT-qPCR in a Taqman assay. M-CSFR fold-change values relative to the S AKYC13 average are indicated on the Y-axis. R-line fish (R) are shown in grey, S-line fish (S) in black. Average +/− SE. *: p

    Journal: Developmental and comparative immunology

    Article Title: Innate Immune Cell Signatures in a BCWD-Resistant Line Of Rainbow Trout Before and After In Vivo Challenge With Flavobacterium psychrophilum

    doi: 10.1016/j.dci.2018.08.018

    Figure Lengend Snippet: Gene expression of M-CSFR in 4 rainbow trout immune tissues (AK, PBL, SPL, and PK) from YC13 and/or YC15, using RT-qPCR in a Taqman assay. M-CSFR fold-change values relative to the S AKYC13 average are indicated on the Y-axis. R-line fish (R) are shown in grey, S-line fish (S) in black. Average +/− SE. *: p

    Article Snippet: Expression in AK, spleen (SPL), peripheral blood leukocytes (PBL) and posterior kidney (PK) from YC13 or YC15 fish were determined using a custom TaqMan® Gene Expression Assay (Applied Biosystems), with a FAM reporter, NFQ quencher, and a ROX reference.

    Techniques: Expressing, Quantitative RT-PCR, TaqMan Assay, Fluorescence In Situ Hybridization