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Real-time quantitative PCR analysis of human colonic HT-29 cells under insulin stimulation

ABSTRACT: Compared mRNA expression between control and insulin treated cells. HT29 cells were treated with 100nM Insulin for 24h. The RT2 Profiler PCR Arrays PAHS-30C (SA Biosciences, MD, USA) was designed to analyze 84 genes related to human insulin signaling pathway. The RT-PCR was carried out using an ABI PRISM® 7000 Sequence Detection System (Applied Biosystems, Foster City, CA).The expression of HSD11B2 for the 3 experiments concerned was monitored in parallel by real time PCR which confirming significant downregulation by insulin.

ORGANISM(S): Homo sapiens

PROVIDER: GSE51677 | GEO | 2014/06/01

SECONDARY ACCESSION(S): PRJNA224313

REPOSITORIES: GEO

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Real-time quantitative PCR analysis of human colonic HT-29 cells under insulin stimulation

Project description:Compared mRNA expression between control and insulin treated cells. HT29 cells were treated with 100nM Insulin for 24h. The RT2 Profiler PCR Arrays PAHS-30C (SA Biosciences, MD, USA) was designed to analyze 84 genes related to human insulin signaling pathway. The RT-PCR was carried out using an ABI PRISMM-BM-. 7000 Sequence Detection System (Applied Biosystems, Foster City, CA).The expression of HSD11B2 for the 3 experiments concerned was monitored in parallel by real time PCR which confirming significant downregulation by insulin. qPCR gene expression profiling. RNA from 3 separated experiments was collected.

2014-06-01 | E-GEOD-51677 | biostudies-arrayexpress

Real-time quantitative analysis of genes related to human innate & adaptive immune responses in neuroendocrine liver metastasis.

Project description:The immunological profile of patient-derived Precision Cut Tissue Slices (PCTS) obtained from neuroendocrine liver metastasis (LM-NEN=liver metastasis of neuroendocrine neoplasm) (n=3), cholangio carcinoma (CCA) (n=1) and tumour-free liver tissue (n=4) was analysed and the expression of 84 genes involved in innate and adaptive immune response was assessed. PCTS were kept in culture at 37C for 2 hours (recovery step) in a sealed chamber filled with carbogen under constant shaking prior to collection. Upon collection tissue slices were preserved in AllProtect Tissue Reagent (Qiagen) and stored at -80C until processing. RNA was isolated with Allprep Mini Kit (Qiagen) according to manifacturer's instructions. cDNA was synthesized with RT2 First Strand Kit (Qiagen) and analysed with Innate and Adaptive Immune Responses RT2 Profiler PCR Array (Qiagen) with RT2 SYBR® Green qPCR Mastermix (Qiagen). qPCR array for Human Innate & Adaptive Immune Responses (Qiagen 330231 PAHS-052ZA, full list of genes available online) was run on ABI 7500 Real-Time PCR System with an initial denaturation step at 95°C for 10 minutes followed by 40 cycles of denaturation at 95°C for 15 seconds and annealing/extension at 60°C for 60 seconds. More information about the histology of the slices used in this analysis can be found in the original manuscipt Doornebal et al.

2022-06-28 | GSE206828 | GEO

Human Stress and Toxicity PathwayFinder PCR Array

Project description:Whole blood was collected for styrene-exposed and control workers subjects in a fiberglass boat industry. Total RNA was extracted from blood lymphocytes, and cDNA was synthesized for profiling on the Human Stress and Toxicity PathwayFinder(tm) RT2 Profiler(tm) PCR Expression Array (PAHS-003, SABiosciences).

2013-09-23 | E-MTAB-1779 | biostudies-arrayexpress

Real-time Quantitative PCR analysis of the expression of TGFB-pathway target genes on 3D-cultured chemoresistant colon carcinoma cells

Project description:The aim of the study was to elucidate which TGFB signaling pathway molecules are involved in the colon cancer chemoresistance. The PCR array used for this experiments was 84 Human TGF-Î² Signaling Targets (Cat. N. PAHS-235ZA, Qiagen) The expression of 84 Human TGF-Î² Signaling Targets genes was analyzed by RT2 profiler PCR array (PAHS-235ZA, Qiagen) using the StepOne Plus instrument (Applied Biosystems) following the manufacturerâs protocol. Two independent experiments were performed for each group of treatment. Untreated cells were used as reference control sample. The mRNA expression levels of each gene in each cell treatment were normalized using the expression of the housekeeping genes B2M, GAPDH, RPLP0, HPRT1 and ACTB.

2016-03-10 | E-GEOD-77927 | biostudies-arrayexpress

Real-time quantitative PCR analysis of UPR genes in human normal and osteoarthritic chondrocytes

Project description:Human chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from individuals within 12 hours of death (normal) and from patients undergoing total knee arthroplasty (osteoarthritic, OA). The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. Total RNA was extracted from the primary chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73749 | GEO

RT2 Profiler PCR array of FoxO1 related genes in mice cardiomyocytes

Project description:mRNA levels of the known 91 FoxO1 target genes were evaluated with RT2 Profiler PCR array in cardiomyocytes transduced with LacZ, Mst1, FoxO1, FoxO1+Mst1, or FoxO1+DN-Mst1 genes were evaluated with RT2 Profiler PCR arrays.

2022-09-12 | GSE213011 | GEO

Real-time quantitative PCR analysis of UPR genes in human osteoarthritic chondrocytes specimens treated with thapsigargin compared to the untreated (control) chondrocytes

Project description:Human osteoarthritic (OA) chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from three patients undergoing total knee arthroplasty. The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. The chondrocytes from each of the three donors were left untreated (control) and treated with thapsigargin (50nM for 20hours).Total RNA was extracted from the chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73746 | GEO

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in systemic sclerosis (SSc) dermal fibroblasts

Project description:Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs.

2012-01-01 | GSE34142 | GEO

Real-time quantitative PCR analysis of UPR genes in human osteoarthritic chondrocytes specimens treated with tunicamycin compared to the untreated (control) chondrocytes

Project description:Human osteoarthritic (OA) chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from three patients undergoing total knee arthroplasty. The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. The chondrocytes from each of the three donors were left untreated (control) and treated with tunicamycin (500ng/ml for 20 hours).Total RNA was extracted from the chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73748 | GEO

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in involved skin of systemic scleroderma (SSc), localized scleroderma (LSc) and Keloid.

Project description:Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. The microRNA (miRNA) isolation from human skin tissue was performed using miRNeasy FFPE kit (Qiagen). For PCR array, miRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of miRNAs from 3 normal skins, 3 SSc, 3 LSc or 3 keloid were prepared, and miRNA expression profile in each disease in vivo was evaluated using RT2 Profiler PCR Array. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into 96-well RT2 miRNA PCR Array that includes primer pairs for 88 human miRNAs (SABiosciences).

2013-01-15 | GSE43469 | GEO

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