trypsin edta  (Lonza)


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    Name:
    Trypsin EDTA
    Description:
    Trypsin EDTA 10X includes 5g L trypsin 1 250 and 2g L Versene EDTA manufactured with irradiated parcine trypsine tested for porcine parvovirus and mycoplasma 100 mL
    Catalog Number:
    BE02-007E
    Price:
    None
    Category:
    Culture Media and Reagents
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    Structured Review

    Lonza trypsin edta
    Trypsin EDTA 10X includes 5g L trypsin 1 250 and 2g L Versene EDTA manufactured with irradiated parcine trypsine tested for porcine parvovirus and mycoplasma 100 mL
    https://www.bioz.com/result/trypsin edta/product/Lonza
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trypsin edta - by Bioz Stars, 2021-05
    96/100 stars

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    Centrifugation:

    Article Title: Phosphorylation of human TRM9L integrates multiple stress-signaling pathways for tumor growth suppression
    Article Snippet: .. Unless otherwise stated, cells were detached from tissue culture plates by trypsinization (0.05% trypsin-EDTA without Ca2+ /Mg2+ , Lonza) at 37°C and collected by centrifugation (500 g , 5 min, 4°C). .. After aspirating medium, the cell pellet was resuspended in lysis buffer consisting of PBS, 1% Triton X-100, 2 mM EDTA, 1:100 mammalian cell protease inhibitors (Sigma-Aldrich), and 1:100 phosphatase inhibitor (Halt, Thermo Fisher Scientific).

    Cell Cycle Assay:

    Article Title: Diatom-Derived Polyunsaturated Aldehydes Activate Cell Death in Human Cancer Cell Lines but Not Normal Cells
    Article Snippet: Densitometric analysis of immunopositive bands was performed using Image J software. .. Cell cycle analysis A549 cells were collected from plates using 1 mL of Trypsin-EDTA (Lonza, Italy), fixed in 70% ethanol and stored at −20°C. .. Cells were then washed twice with PBS, resuspended in PBS containing 1 mg ml−1 RNase A (Qiagen, Cat.19101), incubated at 37°C for 45 min and then stained with propidium iodide (PI, 10 µg ml−1 ) for 15 min.

    Modification:

    Article Title: A microcarrier-based spheroid 3D invasion assay to monitor dynamic cell movement in extracellular matrix
    Article Snippet: In conclusion, this microcarrier-based 3D invasion assay is a powerful tool to study cell invasion in vitro. .. Reagents Dulbecco’s modified Eagle’s medium (DMEM, D0819, Sigma); Trypsin-EDTA (BE-17-161E, Lonza); Dulbecco’s Phosphate Buffered Saline (PBS, Ca2+ and Mg2+ free, D8537, Sigma-Aldrich); Fetal bovine serum (FBS, F7524, Sigma); Collagen type I, rat tail (08–115; Millipore); Matrigel Growth factor reduced (356,231, Coring); Agar (A1296, Sigma-Aldrich); Sodium bicarbonate (11810–017, Life technologies). .. Imaging System and Climate Control Configuration As time-lapse imaging may take hours to days, a screening system, e.g. confocal microscope, integrated with a cell incubation setup is indispensable.

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  • mscs  (Lonza)
    96
    Lonza mscs
    Inflammation enhances the mineralization process in <t>ha-MSCs.</t> a After inflammatory stimulation, ha-MSCs were cultured with specific osteogenic and adipogenic induction media for 21 and 14 days, respectively. b Calcium mineralization process was significantly marked under inflammatory conditions, mainly after <t>AAA-PBMC</t> influence as showed by Alizarin Red staining. Quantification values are represented as mean ± standard deviation and compared with induced ha-MSCs. c MMP-9 detection on osteogenic differentiated ha-MSCs was performed by immunofluorescence, revealing an appreciable staining only after osteogenic induction; 20× magnification. d Oil Red O staining of lipid droplets in ha-MSCs was reduced after priming cells with inflammatory cytokines, as shown by e PPAR-γ mRNA. Results expressed as fold changes relative to induced ha-MSCs. * p
    Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mscs/product/Lonza
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    97
    Lonza g0 by trypsination
    Upregulation of ENDOV transcription during quiescence. ( A ) Human embryonic fibroblasts were arrested in <t>G0</t> by serum starvation at confluence and released by replating 1∶4 in culture medium with serum. ENDOV transcript levels (exons 2 to 3) were measured during cell cycle progression after G0 release at indicated time points by qRT-PCR. G0 cells were used as the reference for calculations. The average of 3 parallels (same RNA) was calculated and standard deviation is shown. Cell cycle distribution was monitored using propidium iodide staining followed by flow cytometry after release from the block. The percentage of cells in each cell cycle is presented in the table. The experiment was repeated twice with similar results. ( B ) Nothern blot analysis of ENDOV mRNA. mRNA was isolated from human fibroblasts that were unsynchronised (u), G0 arrested (G0) and allowed to proliferate for 24 hours (t 24 ), separated by electrophoresis and transferred to a nylon membrane. Hybridisation signals with probes spanning exons 4–8, exon 10, exon 3 of ENDOV and for β -ACTIN are shown. M is the RNA size standard as indicated (in kilobases).
    G0 By Trypsination, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Inflammation enhances the mineralization process in ha-MSCs. a After inflammatory stimulation, ha-MSCs were cultured with specific osteogenic and adipogenic induction media for 21 and 14 days, respectively. b Calcium mineralization process was significantly marked under inflammatory conditions, mainly after AAA-PBMC influence as showed by Alizarin Red staining. Quantification values are represented as mean ± standard deviation and compared with induced ha-MSCs. c MMP-9 detection on osteogenic differentiated ha-MSCs was performed by immunofluorescence, revealing an appreciable staining only after osteogenic induction; 20× magnification. d Oil Red O staining of lipid droplets in ha-MSCs was reduced after priming cells with inflammatory cytokines, as shown by e PPAR-γ mRNA. Results expressed as fold changes relative to induced ha-MSCs. * p

    Journal: Stem Cell Research & Therapy

    Article Title: The crosstalk between vascular MSCs and inflammatory mediators determines the pro-calcific remodelling of human atherosclerotic aneurysm

    doi: 10.1186/s13287-017-0554-x

    Figure Lengend Snippet: Inflammation enhances the mineralization process in ha-MSCs. a After inflammatory stimulation, ha-MSCs were cultured with specific osteogenic and adipogenic induction media for 21 and 14 days, respectively. b Calcium mineralization process was significantly marked under inflammatory conditions, mainly after AAA-PBMC influence as showed by Alizarin Red staining. Quantification values are represented as mean ± standard deviation and compared with induced ha-MSCs. c MMP-9 detection on osteogenic differentiated ha-MSCs was performed by immunofluorescence, revealing an appreciable staining only after osteogenic induction; 20× magnification. d Oil Red O staining of lipid droplets in ha-MSCs was reduced after priming cells with inflammatory cytokines, as shown by e PPAR-γ mRNA. Results expressed as fold changes relative to induced ha-MSCs. * p

    Article Snippet: After monocyte adhesion to plastic flask (4 hours), AAA-PBMCs were counted and plated on an MSC feeder layer at a density ratio of 1:2 MSCs:PBMCs in RPMI 1640 (Lonza); 5 μg/ml phytohaemagglutin (PHA; Sigma Aldrich) was added to activate AAA-PBMCs.

    Techniques: Cell Culture, Staining, Standard Deviation, Immunofluorescence

    ha-MSCs exposed to inflammatory conditions assume a pathological phenotype. a PBMCs isolated from AAA patients showed a molecular signature reporting high levels of inflammatory cytokines (TNF-α and IL-1β) and reduced anti-inflammatory IL-10. Results are expressed as fold changes relative to healthy PBMCs. b According to the experimental design, ha-MSCs were exposed to inflammatory mediators (cytokines and PBMCs) for 24 hours, then investigated in terms of vascular remodelling and differentiation properties. ha-MSCs exposed to inflammation underwent increased transcription of ( c ) MMP-9 and ( d ) osteogenic lineage-specific markers (BMP-2, OPN, OCN), to the detriment of the adipogenic transcriptional factor PPAR-γ. Results are expressed as fold changes relative to unexposed ha-MSCs. * p

    Journal: Stem Cell Research & Therapy

    Article Title: The crosstalk between vascular MSCs and inflammatory mediators determines the pro-calcific remodelling of human atherosclerotic aneurysm

    doi: 10.1186/s13287-017-0554-x

    Figure Lengend Snippet: ha-MSCs exposed to inflammatory conditions assume a pathological phenotype. a PBMCs isolated from AAA patients showed a molecular signature reporting high levels of inflammatory cytokines (TNF-α and IL-1β) and reduced anti-inflammatory IL-10. Results are expressed as fold changes relative to healthy PBMCs. b According to the experimental design, ha-MSCs were exposed to inflammatory mediators (cytokines and PBMCs) for 24 hours, then investigated in terms of vascular remodelling and differentiation properties. ha-MSCs exposed to inflammation underwent increased transcription of ( c ) MMP-9 and ( d ) osteogenic lineage-specific markers (BMP-2, OPN, OCN), to the detriment of the adipogenic transcriptional factor PPAR-γ. Results are expressed as fold changes relative to unexposed ha-MSCs. * p

    Article Snippet: After monocyte adhesion to plastic flask (4 hours), AAA-PBMCs were counted and plated on an MSC feeder layer at a density ratio of 1:2 MSCs:PBMCs in RPMI 1640 (Lonza); 5 μg/ml phytohaemagglutin (PHA; Sigma Aldrich) was added to activate AAA-PBMCs.

    Techniques: Isolation

    VSMC-induced MSC proliferation requires mitochondrial transfer from VSMCs to MSCs. Graph showing MFI represents the cell proliferation in CFSE-labeled MSCs and in cocultures with control VSMCs and with VSMCs having mitochondrial dysfunction. Cell proliferation

    Journal: Stem Cells and Development

    Article Title: Vascular Smooth Muscle Cells Initiate Proliferation of Mesenchymal Stem Cells by Mitochondrial Transfer via Tunneling Nanotubes

    doi: 10.1089/scd.2011.0691

    Figure Lengend Snippet: VSMC-induced MSC proliferation requires mitochondrial transfer from VSMCs to MSCs. Graph showing MFI represents the cell proliferation in CFSE-labeled MSCs and in cocultures with control VSMCs and with VSMCs having mitochondrial dysfunction. Cell proliferation

    Article Snippet: Monocultures and cocultures of MSCs and VSMCs were grown in a medium consisting of an MSC basal medium (MSCBM; Lonza Walkersville, Inc.), supplemented with 2% fetal bovine serum (Promocell GmbH), 4 mM l -glutamine (Lonza Walkersville, Inc.), and 0.01% penicillin/streptomycin (Biochrom AG).

    Techniques: Labeling

    VSMC-induced MSC proliferation requires TNT formation. (a) Graph showing MFI represents the cell proliferation in CFSE-labeled MSCs and VSMC cocultures. Increase in MSC proliferation was completely abolished by TNT disruption with cytochalasin D (1 μM)

    Journal: Stem Cells and Development

    Article Title: Vascular Smooth Muscle Cells Initiate Proliferation of Mesenchymal Stem Cells by Mitochondrial Transfer via Tunneling Nanotubes

    doi: 10.1089/scd.2011.0691

    Figure Lengend Snippet: VSMC-induced MSC proliferation requires TNT formation. (a) Graph showing MFI represents the cell proliferation in CFSE-labeled MSCs and VSMC cocultures. Increase in MSC proliferation was completely abolished by TNT disruption with cytochalasin D (1 μM)

    Article Snippet: Monocultures and cocultures of MSCs and VSMCs were grown in a medium consisting of an MSC basal medium (MSCBM; Lonza Walkersville, Inc.), supplemented with 2% fetal bovine serum (Promocell GmbH), 4 mM l -glutamine (Lonza Walkersville, Inc.), and 0.01% penicillin/streptomycin (Biochrom AG).

    Techniques: Labeling

    MSC coculture with VSMCs does not induce MSC differentiation. (a) Representative flow cytometric analysis shows no change of VSMC markers α-SMA and calponin in MSCs and VSMCs in mono- and cocultures; n =3. (b) Expression of VSMC markers in MSCs

    Journal: Stem Cells and Development

    Article Title: Vascular Smooth Muscle Cells Initiate Proliferation of Mesenchymal Stem Cells by Mitochondrial Transfer via Tunneling Nanotubes

    doi: 10.1089/scd.2011.0691

    Figure Lengend Snippet: MSC coculture with VSMCs does not induce MSC differentiation. (a) Representative flow cytometric analysis shows no change of VSMC markers α-SMA and calponin in MSCs and VSMCs in mono- and cocultures; n =3. (b) Expression of VSMC markers in MSCs

    Article Snippet: Monocultures and cocultures of MSCs and VSMCs were grown in a medium consisting of an MSC basal medium (MSCBM; Lonza Walkersville, Inc.), supplemented with 2% fetal bovine serum (Promocell GmbH), 4 mM l -glutamine (Lonza Walkersville, Inc.), and 0.01% penicillin/streptomycin (Biochrom AG).

    Techniques: Flow Cytometry, Expressing

    MSCs and VSMCs form TNT-like structures for intercellular contacts. Flow cytometry of cocultured MSCs and VSMCs. MSCs were labeled with CellTracker ™ (CM-DiI), whereas VSMCs were unlabeled; cytochalasin D (1 μM) was used to disrupt

    Journal: Stem Cells and Development

    Article Title: Vascular Smooth Muscle Cells Initiate Proliferation of Mesenchymal Stem Cells by Mitochondrial Transfer via Tunneling Nanotubes

    doi: 10.1089/scd.2011.0691

    Figure Lengend Snippet: MSCs and VSMCs form TNT-like structures for intercellular contacts. Flow cytometry of cocultured MSCs and VSMCs. MSCs were labeled with CellTracker ™ (CM-DiI), whereas VSMCs were unlabeled; cytochalasin D (1 μM) was used to disrupt

    Article Snippet: Monocultures and cocultures of MSCs and VSMCs were grown in a medium consisting of an MSC basal medium (MSCBM; Lonza Walkersville, Inc.), supplemented with 2% fetal bovine serum (Promocell GmbH), 4 mM l -glutamine (Lonza Walkersville, Inc.), and 0.01% penicillin/streptomycin (Biochrom AG).

    Techniques: Flow Cytometry, Cytometry, Labeling

    Intercellular exchange of mitochondria between MSCs and VSMCs. MSCs or VSMCs were labeled with either MitoTracker Red ( red ) or CFSE ( green ). MitoTracker Red-labeled cells were cocultured for 2 and 24 h with CFSE-labeled cells. Fluorescence confocal

    Journal: Stem Cells and Development

    Article Title: Vascular Smooth Muscle Cells Initiate Proliferation of Mesenchymal Stem Cells by Mitochondrial Transfer via Tunneling Nanotubes

    doi: 10.1089/scd.2011.0691

    Figure Lengend Snippet: Intercellular exchange of mitochondria between MSCs and VSMCs. MSCs or VSMCs were labeled with either MitoTracker Red ( red ) or CFSE ( green ). MitoTracker Red-labeled cells were cocultured for 2 and 24 h with CFSE-labeled cells. Fluorescence confocal

    Article Snippet: Monocultures and cocultures of MSCs and VSMCs were grown in a medium consisting of an MSC basal medium (MSCBM; Lonza Walkersville, Inc.), supplemented with 2% fetal bovine serum (Promocell GmbH), 4 mM l -glutamine (Lonza Walkersville, Inc.), and 0.01% penicillin/streptomycin (Biochrom AG).

    Techniques: Labeling, Fluorescence

    The expression level of osteocalcin or bone gamma-carboxyglutamic acid-containing protein ( BGLAP ) gene in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of osteocalcin gene expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest expression of osteocalcin gene was detected on day 14. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Journal: Bosnian Journal of Basic Medical Sciences

    Article Title: Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

    doi: 10.17305/bjbms.2018.2633

    Figure Lengend Snippet: The expression level of osteocalcin or bone gamma-carboxyglutamic acid-containing protein ( BGLAP ) gene in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of osteocalcin gene expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest expression of osteocalcin gene was detected on day 14. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Article Snippet: The plasmid bearing pre-miR-210 (vector with the target gene) and two control plasmids, one containing pmaxGFP (an empty vector) and the other containing Scramble (vector with a sequence that does not belong to any known miRNA), were transfected separately into MSCs using a Human MSC Nucleofector kit (Lonza, USA) and U23 program.

    Techniques: Expressing, Transfection, Plasmid Preparation

    Overexpression of miR-210 in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of miR-210 expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210 and MSCs transfected with Scramble on days 7, 14 and 21. The highest expression level of miR-210 was detected on day 7. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Journal: Bosnian Journal of Basic Medical Sciences

    Article Title: Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

    doi: 10.17305/bjbms.2018.2633

    Figure Lengend Snippet: Overexpression of miR-210 in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of miR-210 expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210 and MSCs transfected with Scramble on days 7, 14 and 21. The highest expression level of miR-210 was detected on day 7. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Article Snippet: The plasmid bearing pre-miR-210 (vector with the target gene) and two control plasmids, one containing pmaxGFP (an empty vector) and the other containing Scramble (vector with a sequence that does not belong to any known miRNA), were transfected separately into MSCs using a Human MSC Nucleofector kit (Lonza, USA) and U23 program.

    Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation

    Mesenchymal stem cells (MSCs) were successfully transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210 by electroporation, (A) as observed under optical microscope. (B) Approximately 63% of transfected MSCs emitted green fluorescence under the inverted fluorescence microscope, 48 hours after the transfection. (C) Flow cytometric analysis also confirmed successful transfection of MSCs, i.e., infection rate of 63% was observed.

    Journal: Bosnian Journal of Basic Medical Sciences

    Article Title: Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

    doi: 10.17305/bjbms.2018.2633

    Figure Lengend Snippet: Mesenchymal stem cells (MSCs) were successfully transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210 by electroporation, (A) as observed under optical microscope. (B) Approximately 63% of transfected MSCs emitted green fluorescence under the inverted fluorescence microscope, 48 hours after the transfection. (C) Flow cytometric analysis also confirmed successful transfection of MSCs, i.e., infection rate of 63% was observed.

    Article Snippet: The plasmid bearing pre-miR-210 (vector with the target gene) and two control plasmids, one containing pmaxGFP (an empty vector) and the other containing Scramble (vector with a sequence that does not belong to any known miRNA), were transfected separately into MSCs using a Human MSC Nucleofector kit (Lonza, USA) and U23 program.

    Techniques: Transfection, Plasmid Preparation, Electroporation, Microscopy, Fluorescence, Flow Cytometry, Infection

    The expression level of runt-related transcription factor 2 (Runx2) in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of Runx2 expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest expression of Runx2 was detected on day 7. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Journal: Bosnian Journal of Basic Medical Sciences

    Article Title: Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

    doi: 10.17305/bjbms.2018.2633

    Figure Lengend Snippet: The expression level of runt-related transcription factor 2 (Runx2) in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of Runx2 expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest expression of Runx2 was detected on day 7. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Article Snippet: The plasmid bearing pre-miR-210 (vector with the target gene) and two control plasmids, one containing pmaxGFP (an empty vector) and the other containing Scramble (vector with a sequence that does not belong to any known miRNA), were transfected separately into MSCs using a Human MSC Nucleofector kit (Lonza, USA) and U23 program.

    Techniques: Expressing, Transfection, Plasmid Preparation

    The expression level of alkaline phosphatase (ALP, ALPL ) gene in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of ALP expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest ALP gene expression was detected on day 21. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Journal: Bosnian Journal of Basic Medical Sciences

    Article Title: Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

    doi: 10.17305/bjbms.2018.2633

    Figure Lengend Snippet: The expression level of alkaline phosphatase (ALP, ALPL ) gene in mesenchymal stem cells (MSCs) on different days of transfection. Fold increase ± standard error of the mean (SEM) of ALP expression was compared between MSCs transfected with plenti-III-mir-green fluorescent protein (GFP) plasmid bearing pre-miR-210, MSCs transfected with Scramble and MSCs transfected with pmaxGFP vector on days 0, 7, 14 and 21. The highest ALP gene expression was detected on day 21. The difference in gene expression was calculated using the 2-ΔΔCT method ( p

    Article Snippet: The plasmid bearing pre-miR-210 (vector with the target gene) and two control plasmids, one containing pmaxGFP (an empty vector) and the other containing Scramble (vector with a sequence that does not belong to any known miRNA), were transfected separately into MSCs using a Human MSC Nucleofector kit (Lonza, USA) and U23 program.

    Techniques: Expressing, ALP Assay, Transfection, Plasmid Preparation

    Upregulation of ENDOV transcription during quiescence. ( A ) Human embryonic fibroblasts were arrested in G0 by serum starvation at confluence and released by replating 1∶4 in culture medium with serum. ENDOV transcript levels (exons 2 to 3) were measured during cell cycle progression after G0 release at indicated time points by qRT-PCR. G0 cells were used as the reference for calculations. The average of 3 parallels (same RNA) was calculated and standard deviation is shown. Cell cycle distribution was monitored using propidium iodide staining followed by flow cytometry after release from the block. The percentage of cells in each cell cycle is presented in the table. The experiment was repeated twice with similar results. ( B ) Nothern blot analysis of ENDOV mRNA. mRNA was isolated from human fibroblasts that were unsynchronised (u), G0 arrested (G0) and allowed to proliferate for 24 hours (t 24 ), separated by electrophoresis and transferred to a nylon membrane. Hybridisation signals with probes spanning exons 4–8, exon 10, exon 3 of ENDOV and for β -ACTIN are shown. M is the RNA size standard as indicated (in kilobases).

    Journal: PLoS ONE

    Article Title: The Human Homolog of Escherichia coli Endonuclease V Is a Nucleolar Protein with Affinity for Branched DNA Structures

    doi: 10.1371/journal.pone.0047466

    Figure Lengend Snippet: Upregulation of ENDOV transcription during quiescence. ( A ) Human embryonic fibroblasts were arrested in G0 by serum starvation at confluence and released by replating 1∶4 in culture medium with serum. ENDOV transcript levels (exons 2 to 3) were measured during cell cycle progression after G0 release at indicated time points by qRT-PCR. G0 cells were used as the reference for calculations. The average of 3 parallels (same RNA) was calculated and standard deviation is shown. Cell cycle distribution was monitored using propidium iodide staining followed by flow cytometry after release from the block. The percentage of cells in each cell cycle is presented in the table. The experiment was repeated twice with similar results. ( B ) Nothern blot analysis of ENDOV mRNA. mRNA was isolated from human fibroblasts that were unsynchronised (u), G0 arrested (G0) and allowed to proliferate for 24 hours (t 24 ), separated by electrophoresis and transferred to a nylon membrane. Hybridisation signals with probes spanning exons 4–8, exon 10, exon 3 of ENDOV and for β -ACTIN are shown. M is the RNA size standard as indicated (in kilobases).

    Article Snippet: Cell cycle synchronization and analysis by flow cytometrySynchronization of the cells in G0 phase was achieved by culturing cells as a confluent layer for 72 h followed by serum starvation (0.2% serum) for 72 h. The cells were released from G0 by trypsination (Trypsin-EDTA 200 mg/l, Lonza) for 4 min at 37°C and cultivated in standard growth medium at 25% confluence.

    Techniques: Quantitative RT-PCR, Standard Deviation, Staining, Flow Cytometry, Blocking Assay, Isolation, Electrophoresis, Hybridization