hmec 1 cells  (Qiagen)


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    Structured Review

    Qiagen hmec 1 cells
    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved <t>HMEC-1</t> cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

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    Images

    1) Product Images from "Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion"

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24659

    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P
    Figure Legend Snippet: Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

    Techniques Used: Incubation, Expressing, Western Blot

    Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P
    Figure Legend Snippet: Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P

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

    Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P
    Figure Legend Snippet: Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P

    Techniques Used: Expressing, Western Blot, Concentration Assay

    Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P
    Figure Legend Snippet: Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P

    Techniques Used: Western Blot, Concentration Assay

    Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P
    Figure Legend Snippet: Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P

    Techniques Used: Cell Culture, Concentration Assay

    Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P
    Figure Legend Snippet: Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P

    Techniques Used: Activation Assay, Stable Transfection, Transfection, Luciferase, Concentration Assay, Activity Assay, Incubation

    2) Product Images from "Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion"

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24659

    Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation (** P < 0.01 vs. basal, *** P < 0.001 vs. basal). When pre-incubated with BAY 11-7085 [(10μM), NF-ĸB inhibitor] or U0126 [(10μM), MAPK inhibitor] or SB202190 [(1 μM), p38 MAPK inhibitor] or [(10 μM), PI3K/Akt inhibitor] for 1 hour, chemerin induced (10nM) NF-B activation was significantly attenuated (# P < 0.001 vs. chemerin (10nM) only treated). When chemerin (10nM) was co-incubated with IL-1β (0-100ng/mL), a significant increase in NF-B activity was observed [Figure 1: a P < 0.05, b P < 0.01, c P < 0.001 vs. IL-1β (100ng/mL) only treated, P < 0.001 vs. chemerin (10nM) only treated]. Data are mean ± SE of three experiments. Each experiment was carried out in three replicates. Group comparison by ANOVA (post hoc analysis: Tukey's test).
    Figure Legend Snippet: Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation (** P < 0.01 vs. basal, *** P < 0.001 vs. basal). When pre-incubated with BAY 11-7085 [(10μM), NF-ĸB inhibitor] or U0126 [(10μM), MAPK inhibitor] or SB202190 [(1 μM), p38 MAPK inhibitor] or [(10 μM), PI3K/Akt inhibitor] for 1 hour, chemerin induced (10nM) NF-B activation was significantly attenuated (# P < 0.001 vs. chemerin (10nM) only treated). When chemerin (10nM) was co-incubated with IL-1β (0-100ng/mL), a significant increase in NF-B activity was observed [Figure 1: a P < 0.05, b P < 0.01, c P < 0.001 vs. IL-1β (100ng/mL) only treated, P < 0.001 vs. chemerin (10nM) only treated]. Data are mean ± SE of three experiments. Each experiment was carried out in three replicates. Group comparison by ANOVA (post hoc analysis: Tukey's test).

    Techniques Used: Activation Assay, Stable Transfection, Transfection, Luciferase, Concentration Assay, Activity Assay, Incubation

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    Centrifugation:

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    Amplification:

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    Synthesized:

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    Cytometry:

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    Quantitative RT-PCR:

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    Real-time Polymerase Chain Reaction:

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    Microarray:

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    Incubation:

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    Luciferase:

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    Activity Assay:

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    Derivative Assay:

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    Transfection:

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    Northern Blot:

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    Cell Culture:

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    Reverse Transcription Polymerase Chain Reaction:

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    Article Snippet: Paragraph title: Reverse transcription polymerase chain reaction (RT-PCR) ... Isolation of total RNA, and their subsequent first-strand cDNA synthesis, wasperformed using an RNeasy mini kit (Qiagen, Valencia, CA) and an Omniscript Reverse Transcription kit (Qiagen, Valencia, CA), respectively, per the instructions of manufacturer and as described earlier.

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Paragraph title: Total RNA preparation, RT-PCR, and 5′RACE (5′ rapid amplification of cDNA ends) analysis. ... Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen).

    Generated:

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. Five housekeeping genes (HKGs) and proprietary controls were tested in parallel for normalization of the assay results, monitoring genomic DNA contamination, cDNA synthesis and real-time PCR efficiency.

    DNA Sequencing:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Sequencing:

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Injection:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Recombinant:

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. .. The synthesis of cDNA was performed using the SuperScript First Strand Synthesis System for RT-PCR kit (Invitrogen).

    Fluorescence:

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Mutagenesis:

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: After a two-day growth on TSBYE plates, one colony of the wild-type strain (VT1169) and the rmlC mutant were transferred into 8 ml of freshly prepared TSBYE broth, with or without spectinomycin, and grown overnight. .. Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA).

    Isolation:

    Article Title: Triheptanoin: long-term effects in the very long-chain acyl-CoA dehydrogenase-deficient mouse
    Article Snippet: Signals were detected and quantified Fusion FX Analyzer (Peqlab, Erlangen, Germany). .. Total RNA from heart was isolated with the RNeasy mini kit (Qiagen, Hilden, Germany). .. Forward and reverse primers were designed with the Primer Design Tool from NCBI ( ).

    Article Title: CXCR3/Ligands Are Significantly Involved in the Tumorigenesis of Basal Cell Carcinomas
    Article Snippet: Samples were removed from the stabilization reagent, and homogenized using a QIAshredder kit (Qiagen) according to the manufacturer’s instructions for total RNA isolation. .. Total RNA of the lysates was isolated using the RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. .. DNase digestion was conducted using RNase-Free DNase Set (Qiagen) after the first buffer washing step of the RNA-bound RNeasy membrane.

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: After stripping of phospho-αAMP kinase blots, total AMP kinase (Cell Signaling Technology) immunoreactivity was determined. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. For primary hepatocytes, cells were washed 2× with PBS and lysed with 500 μl of RNeasy lysis buffer supplied with the kit.

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: Paragraph title: Isolation of total RNA from A. actinomycetemcomitans ... Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA).

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: To follow the course of embryonic development, a time-course analysis for multilineage differentiation was performed. hESCs were randomly differentiated as described in , and samples were collected for day 0, 3, 6, 9, 12, 15, 18 and 21 for RT-PCR analysis. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Primer design was performed using Primer3 ( ).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: The number of passages of the RAC and SAC were closely matched for the populations derived from the 13M (passage 7 for RAC and passage 6 for SAC), 57M (passage 6 for both populations), and 70M donor tissues (passage 3 for both populations). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA). .. Gene expression was measured using real-time quantitative PCR (see Supplementary Materials and Methods for detailed methods).

    Article Title: Inhibition of Histone Lysine Methylation Enhances Expression of Cancer-Testis Genes in Lung Cancer Cells: Implications for Adoptive Immunotherapy of Cancer
    Article Snippet: DAC/DP and DAC/TSA treatments were performed as described ( ). .. RNA was isolated using RNeasy mini kit (Qiagen, Valencia, CA). cDNAs were made using reverse transcription kit (Bio-Rad; Hercules, CA). qRT-PCR primers for CT-X genes and β-actin expression are listed in . .. Total cell proteins were extracted and immunoblotting was performed as described ( ) with minor modifications, using primary antibodies listed in , appropriate horseradish peroxidase–conjugated secondary antibodies, and SuperSignal West Pico Chemiluminescent Substrate (Pierce Biotechnology, Rockford, IL).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively.

    Article Title: Reprogramming of skeletal myoblasts for induction of pluripotency for tumor free cardiomyogenesis in the infarcted heart
    Article Snippet: The murine SiPS clone Raf1 was expanded on mitotically inactivated murine embryonic fibroblasts (MEFs; 5×104 cells/cm2 ) as described in . .. Isolation of total RNA, and their subsequent first-strand cDNA synthesis, wasperformed using an RNeasy mini kit (Qiagen, Valencia, CA) and an Omniscript Reverse Transcription kit (Qiagen, Valencia, CA), respectively, per the instructions of manufacturer and as described earlier. .. The primer sequences used are given in .

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: The expression of cytokines involved in Th1 and Th2 responses was evaluated in the lung tissue of immunized mice post-challenge. .. Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. A mouse Th1 and Th2 RT2 -PCR Profiler Array (Qiagen, PAMM-034Z) was used to measure the expression level of 84 T-helper-associated genes in the ABI Prism 7500 Sequence Detection System (ThermoFisher).

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Sf9 cells were infected with AcMNPV at a multiplicity of infection (MOI) of 10 TCID50 . .. Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen). .. Reverse transcription-PCR (RT-PCR) was performed with an RNA PCR kit (v3.0; TaKaRa) using 2.0 μg of total RNA as the template for each time point.

    Subcloning:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Negative Control:

    Article Title:
    Article Snippet: Small interfering RNAs for cathepsin L (siCatL), cathepsin S (siCatS), cathepsin B (siCatB), and a negative control siRNA (siCTL) were obtained from Qiagen SA (Courtaboeuf, France) ( ). .. Total RNAs were extracted at different times (24, 48, 72, and 96 h) (RNeasy Mini Kit, Qiagen SA), and reverse transcription was performed on total RNA (1 mg) using RevertAid M-MuLV Reverse Transcriptase (GmbH, Germany).

    Flow Cytometry:

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Labeling:

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Mouse Assay:

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: VNOs were obtained from Bcl11b−/− and wild-type mice at P0 and stored in RNA later (Ambion, Austin, TX). .. The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany).

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: The expression of cytokines involved in Th1 and Th2 responses was evaluated in the lung tissue of immunized mice post-challenge. .. Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401).

    Polymerase Chain Reaction:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit.

    Article Title: An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers
    Article Snippet: Paragraph title: Semiquantitative PCR of WT CPE and CPE-ΔN transcripts in HCC cells. ... RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science).

    Article Title: Profilin-1 overexpression inhibits proliferation of MDA-MB-231 breast cancer cells partly through p27kip1 upregulation
    Article Snippet: Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA). .. Total RNA was extracted from cells cultured in normal serum-containing growth medium using the RNeasy mini kit commercially available through Qiagen (Valencia, CA).

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: To follow the course of embryonic development, a time-course analysis for multilineage differentiation was performed. hESCs were randomly differentiated as described in , and samples were collected for day 0, 3, 6, 9, 12, 15, 18 and 21 for RT-PCR analysis. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Primer design was performed using Primer3 ( ).

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells
    Article Snippet: mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. .. mIMCD-K2 cells were grown to resistance on permeable supports, and total RNA was harvested using an RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions.

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title:
    Article Snippet: Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions. .. Real time RT-PCR was performed on 384-well plates in a 20-μl reaction system containing 2 μl of diluted cDNA, 1 μl of primer mixture, 7 μl of H2 O, and 10 μl of TaqMan 2× in reaction mixture.

    Construct:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Purification:

    Article Title: Lipopolysaccharides mediate leukotoxin secretion in Aggregatibacter actinomycetemcomitans
    Article Snippet: The cells were centrifuged at 5000 ×g for 10 min and resuspended in 100 µl TE buffer (10 mM Tris-Cl, 1 mM EDTA, pH 8.0) containing 1 mg/ml of lysozyme Type VI (MP Biomedicals, Aurora, OH). .. Total RNA was purified from the lysate using an RNeasy Mini Kit (QIAGEN Inc., Valencia, CA). .. The purified total RNA was pretreated with deoxyribonuclease I (DNase), amplification grade (Invitrogen, Carlsbad, CA), to remove contaminating DNA.

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: RNA was purified from 1 × 105 cells for gene expression analysis immediately following the thaw of the cryopreserved stocks of cultured cells. .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Article Title: Identification of Autographa californica Nucleopolyhedrovirus ac93 as a Core Gene and Its Requirement for Intranuclear Microvesicle Formation and Nuclear Egress of Nucleocapsids
    Article Snippet: Total cellular RNA was isolated using an RNeasy minikit (Qiagen), according to the manufacturer's instructions, at various time points postinfection (p.i.) and digested using an RNase-free DNase set (Qiagen). .. The ac93 -specific primers Ac93P1 (5′-ATGGCGACTAGCAAAACGATCG-3′) and Ac93P2 (5′-TTAATTTACAATTTCAATTCC-3′) were used for PCR amplification to detect the transcription of ac93 .

    Chromatin Immunoprecipitation:

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was isolated from synaptosomal homogenates by addition of 10% BCP and standard phase separation, followed by overnight precipitation with isopropanol at -20°C. .. RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Only samples with an RNA integrity number > 8 were included in subsequent analyses. cDNA was synthesized from purified RNA with the ABI High capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA).

    Plasmid Preparation:

    Article Title: Role of oxidative stress in intermittent hypoxia-induced immediate early gene activation in rat PC12 cells
    Article Snippet: Total cellular RNA was isolated using the Rneasy mini kit method (Qiagen Inc., CA, USA) following the manufacturer's protocol. c- fos, TH and β-actin mRNAs were analysed by Northern blot analysis as previously described ( ; ). .. The following plasmids were used: p fos -Luc, wild-type and c- fos promoter with point mutations at Calcium–calcium responsive element (Ca/CRE), Serum responsive element (SRE), and FAP cis -elements with luciferase reporter gene ( ).

    Software:

    Article Title: Bcl11b/Ctip2 controls the differentiation of vomeronasal sensory neurons in mice
    Article Snippet: The total RNA of each preparation was extracted using the RNeasy Mini Kit (Qiagen, Hilden, Germany). .. To ensure reproducibility, microarray analyses were performed with RNA samples from six Bcl11b−/− and five wild-type mice.

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen).

    Article Title: Hippocampal dysregulation of synaptic plasticity-associated proteins with age-related cognitive decline
    Article Snippet: RNA was purified using the Qiagen RNeasy Mini kit (Qiagen), and quality and quantity were assessed by microfluidics chip (Agilent 2100 Expert Bioanalyzer Nano Chip, Agilent, Palo Alto, CA) and spectrometry (NanoDrop ND1000; Thermo Scientific, Wilmington, DE), respectively. .. Gene expression assays included Mtap2 (MAP2, Rn00565046_m1), Dbn1 (drebrin, Rn00578869_m1), Rtn4 (Nogo-A, Rn00582903_m1), Dlgh4 (PSD-95, Rn00571479_m1), Ywhaq (14-3-3θ, Rn00820722_g1) and Camk2 (CaMKIIα, Rn00563883_m1).

    Article Title: Novel recombinant DNA vaccine candidates for human respiratory syncytial virus: Preclinical evaluation of immunogenicity and protection efficiency
    Article Snippet: Lungs were homogenized in liquid nitrogen, and RNA was isolated using an RNeasy Mini Kit (Qiagen, 74104). cDNA synthesis and genomic DNA elimination were achieved using an RT first strand synthesis kit (Qiagen, 330401). .. Five housekeeping genes (HKGs) and proprietary controls were tested in parallel for normalization of the assay results, monitoring genomic DNA contamination, cDNA synthesis and real-time PCR efficiency.

    Electrophoresis:

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. A thermocycler (Bio-Rad, Munich, Germany) was used for PCR reaction, with the following conditions: denaturation at 95°C/2 min, 22 cycles for β-ACTIN and 35 cycles for target genes, of 95°C/30 s denaturation, 60°C/30 s annealing 72°C/60 s of elongation.

    RNA Extraction:

    Article Title:
    Article Snippet: Paragraph title: RNA Extraction, cDNA Synthesis, and Quantitative Real Time PCR Assay ... Total RNA was extracted from IFN-γ-treated or untreated BxPC-3 cells using the RNeasy Mini Kit (catalog number 74104 from Qiagen) following the manufacturer's instructions.

    Article Title: Activation of p38 mitogen-activated protein kinase by norepinephrine in T-lineage cells
    Article Snippet: Paragraph title: RNA extraction and quantitative real-time polymerase chain reaction ... Total cellular RNA was extracted using the RNEasy Mini Prep kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions.

    shRNA:

    Article Title: SIRT4 Regulates Fatty Acid Oxidation and Mitochondrial Gene Expression in Liver and Muscle Cells
    Article Snippet: After stripping of phospho-αAMP kinase blots, total AMP kinase (Cell Signaling Technology) immunoreactivity was determined. .. Total RNA was isolated from primary hepatocytes or primary myotubes transduced with control shRNA and shRNA using the Qiagen RNeasy mini kit. .. For primary hepatocytes, cells were washed 2× with PBS and lysed with 500 μl of RNeasy lysis buffer supplied with the kit.

    Concentration Assay:

    Article Title: Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation
    Article Snippet: Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR. .. Total RNA was isolated from mentioned samples using RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. cDNA synthesis was performed with Super Script Vilo (Invitrogen, Carlsbad, CA, USA) cDNA synthesis kit using 1 µg of total RNA as starting material. cDNA was diluted 10 times with nuclease-free water, and 2 µL was used as a template for PCR.

    Staining:

    Article Title: Interleukin 1? Sustains the Expression of Inflammatory Factors in Human Pancreatic Cancer Microenvironment by Targeting Cancer-Associated Fibroblasts
    Article Snippet: After culture, the cells were harvested, sorted into pure population of PDACs and CAFs, and RNA was extracted using the RNeasy Mini Kit according to the manufacturer's description (Qiagen). .. The double-stranded cDNA was purified using the Sample Cleanup Module and cDNA Cleanup Spin Columns (Affymetrix) and biotin-labeled antisense complementary RNA was synthesized using GeneChip IVT labeling kit (Affymetrix).

    Article Title: Human Skeletal Muscle Cells With a Slow Adhesion Rate After Isolation and an Enhanced Stress Resistance Improve Function of Ischemic Hearts
    Article Snippet: Post-thaw recovery and viability were evaluated from aliquots of the frozen cell suspensions that were prepared and cryopreserved for injection using the Viacount viability stain (Millipore, Hayward, CA) and Guava PCA flow cytometry system (Millipore). .. Total RNA was isolated using the RNeasy Mini kit (Qiagen, Valencia, CA) and was reverse transcribed to cDNA (Applied Biosystems, Foster City, CA).

    Variant Assay:

    Article Title: An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers
    Article Snippet: RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science). .. RNA was extracted from MHCC97L and MHCC97H using the RNeasy Mini Kit (QIAGEN), and first-strand cDNA was synthesized with 1 μg of total RNA from these cells using Transcriptor First Strand cDNA Synthesis Kit (Roche Applied Science).

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    Qiagen hmec 1 cells
    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved <t>HMEC-1</t> cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P
    Hmec 1 Cells, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin and IL-1β co-incubation; synergistic increase in cell adhesion molecule expression Serum starved HMEC-1 cells were treated with chemerin (3nM) with or without IL-1β (10ng/ml) for 12 hours. ( A ) Representative western blots of E-selectin, VCAM-1 and ICAM-1 and their respective β-actin. ( B-D ) Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were synergistically increased when compared to chemerin or IL-1β, alone (Figures 5A-5D: *** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Incubation, Expressing, Western Blot

    Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules mRNA expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 4 hours. Real-time quantitative RT-PCR analyses showed that mRNA expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly up-regulated by chemerin in a concentration dependent manner at 4 hours (Figures 2A-2C : *** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Expressing, Quantitative RT-PCR, Concentration Assay

    Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules protein expression in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 and 24 hours. Densitometric analysis of western blots (cell protein lysates) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that protein expression of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly increased by chemerin in a concentration dependent manner at 12 hours (Figures 3A-3C : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Expressing, Western Blot, Concentration Assay

    Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin increases endothelial cell adhesion molecules protein secretion in HMEC-1 cells Serum starved HMEC-1 cells were treated with chemerin (0-10nM) for 12 hours. Densitometric analysis of western blots (conditioned media) of E-selectin, VCAM-1 and ICAM-1 immune complexes having normalized to β-actin, respectively, showed that secretion of cell adhesion molecules, i.e. E-selectin, VCAM-1 and ICAM-1, were significantly elevated by chemerin in a concentration dependent manner at 12 hours (Figures 4A-4C : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Western Blot, Concentration Assay

    Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin stimulates monocyte-endothelial cell adhesion via NF-ĸB, MAPK and PI3K/Akt pathways Serum starved chemerin (0-10nM) treated HMEC-1 cells were co-cultured with THP-1 cells for 2 hours. Chemerin enhanced monocyte-endothelial adhesion in a concentration dependent fashion (Figure 6A : * P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Cell Culture, Concentration Assay

    Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation ( ** P

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Activation Assay, Stable Transfection, Transfection, Luciferase, Concentration Assay, Activity Assay, Incubation

    Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation (** P < 0.01 vs. basal, *** P < 0.001 vs. basal). When pre-incubated with BAY 11-7085 [(10μM), NF-ĸB inhibitor] or U0126 [(10μM), MAPK inhibitor] or SB202190 [(1 μM), p38 MAPK inhibitor] or [(10 μM), PI3K/Akt inhibitor] for 1 hour, chemerin induced (10nM) NF-B activation was significantly attenuated (# P < 0.001 vs. chemerin (10nM) only treated). When chemerin (10nM) was co-incubated with IL-1β (0-100ng/mL), a significant increase in NF-B activity was observed [Figure 1: a P < 0.05, b P < 0.01, c P < 0.001 vs. IL-1β (100ng/mL) only treated, P < 0.001 vs. chemerin (10nM) only treated]. Data are mean ± SE of three experiments. Each experiment was carried out in three replicates. Group comparison by ANOVA (post hoc analysis: Tukey's test).

    Journal: Oncotarget

    Article Title: Chemerin induces endothelial cell inflammation: activation of nuclear factor-kappa beta and monocyte-endothelial adhesion

    doi: 10.18632/oncotarget.24659

    Figure Lengend Snippet: Chemerin activates NF-кB in HMEC-1 cells via MAPK and PI3K/Akt pathways; synergistic activation with IL-1β pathways Serum starved HMEC-1 cells stably transfected with pNFкB-Luciferase were treated with or without chemerin (0-10nM) for 2 hours. Cells were lysed and luciferase activities were measured. Chemerin induced a concentration dependent increase in luciferase activity after 2 hours of incubation (** P < 0.01 vs. basal, *** P < 0.001 vs. basal). When pre-incubated with BAY 11-7085 [(10μM), NF-ĸB inhibitor] or U0126 [(10μM), MAPK inhibitor] or SB202190 [(1 μM), p38 MAPK inhibitor] or [(10 μM), PI3K/Akt inhibitor] for 1 hour, chemerin induced (10nM) NF-B activation was significantly attenuated (# P < 0.001 vs. chemerin (10nM) only treated). When chemerin (10nM) was co-incubated with IL-1β (0-100ng/mL), a significant increase in NF-B activity was observed [Figure 1: a P < 0.05, b P < 0.01, c P < 0.001 vs. IL-1β (100ng/mL) only treated, P < 0.001 vs. chemerin (10nM) only treated]. Data are mean ± SE of three experiments. Each experiment was carried out in three replicates. Group comparison by ANOVA (post hoc analysis: Tukey's test).

    Article Snippet: Total RNA was extracted from HMEC-1 cells Qiagen RNeasy Mini Kit according to the manufacturer's guidelines (Qiagen, Crawley, UK).

    Techniques: Activation Assay, Stable Transfection, Transfection, Luciferase, Concentration Assay, Activity Assay, Incubation