if-driven sb 10 Search Results


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  • 96
    Millipore 2sb 3ct
    2sb 3ct, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    InvivoGen ncoi ecori digested pdrive meif4a1 vector
    Ncoi Ecori Digested Pdrive Meif4a1 Vector, supplied by InvivoGen, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    InvivoGen mouse initiation factor promoter 4a1 eif sb10
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    Mouse Initiation Factor Promoter 4a1 Eif Sb10, supplied by InvivoGen, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Dojindo Labs fura2 am
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    Fura2 Am, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 92/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen plasmid isolation kit
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    Plasmid Isolation Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 805 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Hamamatsu argus hisca imaging system
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    Argus Hisca Imaging System, supplied by Hamamatsu, used in various techniques. Bioz Stars score: 88/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Promega t7 driven pgem 3z transcription vector
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    T7 Driven Pgem 3z Transcription Vector, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa cytomegalovirus
    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with <t>pT2-IHK-β-gene//eIF-SB10</t> or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples
    Cytomegalovirus, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 469 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa red fluorescent protein dsred2
    Transient expression of red fluorescent protein <t>DsRed2</t> by BME26 cells transfected with a cis- plasmid containing the Sleeping Beauty SB10 transposase and a transposon containing the DsRed2 gene. Data points are the means±standard deviations. Inset:
    Red Fluorescent Protein Dsred2, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Roche fibronectin coated
    Transient expression of red fluorescent protein <t>DsRed2</t> by BME26 cells transfected with a cis- plasmid containing the Sleeping Beauty SB10 transposase and a transposon containing the DsRed2 gene. Data points are the means±standard deviations. Inset:
    Fibronectin Coated, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore p38 stress activated protein kinase sapk inhibitor sb202190
    Response of hepcidin transcript levels to serum deprivation and protein kinase inhibition. ( A ) Serum withdrawal rapidly decreased hepcidin transcript levels in Huh7 cells. ( B ) After 40 h of FCS reduction from 10% to 0.4%, hepcidin transcripts were hardly detectable by RPA in HepG2 cells, whereas hypoxic IGFBP1 induction was not affected. Representative of three independent experiments. U6sn RNA served as loading control. ( C ) Exposure of Huh7 cells to protein kinase inhibitors revealed that the pan kinase inhibitor staurosporine (stauro; 0.5 µM) and the PI3 kinase inhibitor LY294002 (LY; 10 µM) reduced hepcidin expression similar to serum deprivation, whereas the <t>p38</t> SAP kinase inhibitor <t>SB202190</t> (SB; 10 µM) had no effect and the MEK1/2 inhibitor UO126 (1 µM) even increased HAMP/18S ratios. Data are results of qRT PCR analyses and given as means of three independent experiments±SEM; *p
    P38 Stress Activated Protein Kinase Sapk Inhibitor Sb202190, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore m picrotoxin
    Response of hepcidin transcript levels to serum deprivation and protein kinase inhibition. ( A ) Serum withdrawal rapidly decreased hepcidin transcript levels in Huh7 cells. ( B ) After 40 h of FCS reduction from 10% to 0.4%, hepcidin transcripts were hardly detectable by RPA in HepG2 cells, whereas hypoxic IGFBP1 induction was not affected. Representative of three independent experiments. U6sn RNA served as loading control. ( C ) Exposure of Huh7 cells to protein kinase inhibitors revealed that the pan kinase inhibitor staurosporine (stauro; 0.5 µM) and the PI3 kinase inhibitor LY294002 (LY; 10 µM) reduced hepcidin expression similar to serum deprivation, whereas the <t>p38</t> SAP kinase inhibitor <t>SB202190</t> (SB; 10 µM) had no effect and the MEK1/2 inhibitor UO126 (1 µM) even increased HAMP/18S ratios. Data are results of qRT PCR analyses and given as means of three independent experiments±SEM; *p
    M Picrotoxin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 241 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher proton sequencer supplementary
    Response of hepcidin transcript levels to serum deprivation and protein kinase inhibition. ( A ) Serum withdrawal rapidly decreased hepcidin transcript levels in Huh7 cells. ( B ) After 40 h of FCS reduction from 10% to 0.4%, hepcidin transcripts were hardly detectable by RPA in HepG2 cells, whereas hypoxic IGFBP1 induction was not affected. Representative of three independent experiments. U6sn RNA served as loading control. ( C ) Exposure of Huh7 cells to protein kinase inhibitors revealed that the pan kinase inhibitor staurosporine (stauro; 0.5 µM) and the PI3 kinase inhibitor LY294002 (LY; 10 µM) reduced hepcidin expression similar to serum deprivation, whereas the <t>p38</t> SAP kinase inhibitor <t>SB202190</t> (SB; 10 µM) had no effect and the MEK1/2 inhibitor UO126 (1 µM) even increased HAMP/18S ratios. Data are results of qRT PCR analyses and given as means of three independent experiments±SEM; *p
    Proton Sequencer Supplementary, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Tocris sb431542
    <t>SB431542</t> exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p
    Sb431542, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 1892 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Syntaxin syntaxin sx
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    Syntaxin Sx, supplied by Syntaxin, used in various techniques. Bioz Stars score: 88/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore a protinin
    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), <t>syntaxin</t> (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain
    A Protinin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore fibronectin
    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on <t>fibronectin-coated</t> substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P
    Fibronectin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    TaKaRa egfp sequence
    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on <t>fibronectin-coated</t> substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P
    Egfp Sequence, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 599 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher β mercaptoethanol
    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on <t>fibronectin-coated</t> substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P
    β Mercaptoethanol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 18648 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher atg deficient frt hygro fusion gene
    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on <t>fibronectin-coated</t> substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P
    Atg Deficient Frt Hygro Fusion Gene, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Exelixis cher273 tm6 tb
    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on <t>fibronectin-coated</t> substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P
    Cher273 Tm6 Tb, supplied by Exelixis, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Roche fibronectin
    Inactivation of Evl in unconfined MV D7 cells diminishes cell contractile energy. ( A ) Quantifications of contractile energy normalized by cellular area and measured for MV D7 , MVE-KO and Evl-rescue of MVE-KO cells plated on plain polyacrylamide micropatterns coated with <t>fibronectin.</t> ***p≤0.001 or n.s., not significant by Kruskal-Wallis test and Dunn’s Multiple Comparison test. n, number of cells analyzed. Boxes in box plots indicate 50% (25–75%) and whiskers (5–95%) of all measurements, with dashed black lines depicting the medians, arithmetic means are highlighted in red. ( B ) Images depicting representative traction force field representations from cells of distinct genotypes, as indicated. Force scale bar is in Pascal and arrows represent local force magnitude and orientation. Source data for details of contractile energies Figure 8—figure supplement 1 .
    Fibronectin, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 1351 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mbb
    Inactivation of Evl in unconfined MV D7 cells diminishes cell contractile energy. ( A ) Quantifications of contractile energy normalized by cellular area and measured for MV D7 , MVE-KO and Evl-rescue of MVE-KO cells plated on plain polyacrylamide micropatterns coated with <t>fibronectin.</t> ***p≤0.001 or n.s., not significant by Kruskal-Wallis test and Dunn’s Multiple Comparison test. n, number of cells analyzed. Boxes in box plots indicate 50% (25–75%) and whiskers (5–95%) of all measurements, with dashed black lines depicting the medians, arithmetic means are highlighted in red. ( B ) Images depicting representative traction force field representations from cells of distinct genotypes, as indicated. Force scale bar is in Pascal and arrows represent local force magnitude and orientation. Source data for details of contractile energies Figure 8—figure supplement 1 .
    Mbb, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore millex gn syringe driven filter
    Inactivation of Evl in unconfined MV D7 cells diminishes cell contractile energy. ( A ) Quantifications of contractile energy normalized by cellular area and measured for MV D7 , MVE-KO and Evl-rescue of MVE-KO cells plated on plain polyacrylamide micropatterns coated with <t>fibronectin.</t> ***p≤0.001 or n.s., not significant by Kruskal-Wallis test and Dunn’s Multiple Comparison test. n, number of cells analyzed. Boxes in box plots indicate 50% (25–75%) and whiskers (5–95%) of all measurements, with dashed black lines depicting the medians, arithmetic means are highlighted in red. ( B ) Images depicting representative traction force field representations from cells of distinct genotypes, as indicated. Force scale bar is in Pascal and arrows represent local force magnitude and orientation. Source data for details of contractile energies Figure 8—figure supplement 1 .
    Millex Gn Syringe Driven Filter, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with pT2-IHK-β-gene//eIF-SB10 or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples

    Journal: Biochemistry

    Article Title: Erythroid-specific Expression of ?-globin by Sleeping Beauty Transposon for Sickle Cell Disease

    doi: 10.1021/bi6024484

    Figure Lengend Snippet: SB 10 mediated β-globin transposition and identification of the insertion sites in K-562 cells transfected with pT2-IHK-β-gene//eIF-SB10 or pT2-IHK-β-gene. (A) Schematic design for inverted nested PCR analysis. The genomic DNA samples

    Article Snippet: The cis pT2 vectors contained the SB10 transposase gene driven by mouse initiation factor promoter 4A1 (eIF-SB10) (InvivoGen, San Diego, CA).

    Techniques: Transfection, Nested PCR

    Identification of the insertion sites in K-562 cells transfected with pT2-IHK-β-gene//eIF-SB10. The amplicons resulting from the second inverted PCR reactions were isolated from the agarose gel and sequenced directly using the RP2 or LP2 primer

    Journal: Biochemistry

    Article Title: Erythroid-specific Expression of ?-globin by Sleeping Beauty Transposon for Sickle Cell Disease

    doi: 10.1021/bi6024484

    Figure Lengend Snippet: Identification of the insertion sites in K-562 cells transfected with pT2-IHK-β-gene//eIF-SB10. The amplicons resulting from the second inverted PCR reactions were isolated from the agarose gel and sequenced directly using the RP2 or LP2 primer

    Article Snippet: The cis pT2 vectors contained the SB10 transposase gene driven by mouse initiation factor promoter 4A1 (eIF-SB10) (InvivoGen, San Diego, CA).

    Techniques: Transfection, Polymerase Chain Reaction, Isolation, Agarose Gel Electrophoresis

    Transient expression of red fluorescent protein DsRed2 by BME26 cells transfected with a cis- plasmid containing the Sleeping Beauty SB10 transposase and a transposon containing the DsRed2 gene. Data points are the means±standard deviations. Inset:

    Journal: Insect biochemistry and molecular biology

    Article Title: Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

    doi: 10.1016/j.ibmb.2008.01.006

    Figure Lengend Snippet: Transient expression of red fluorescent protein DsRed2 by BME26 cells transfected with a cis- plasmid containing the Sleeping Beauty SB10 transposase and a transposon containing the DsRed2 gene. Data points are the means±standard deviations. Inset:

    Article Snippet: In this construct the SB 10 transposase gene, driven by the cytomegalovirus (CMV) promoter, resided outside the IR/DR of the transposable element containing the reporter gene, red fluorescent protein DsRed2 (Clontech).

    Techniques: Expressing, Transfection, Plasmid Preparation

    Response of hepcidin transcript levels to serum deprivation and protein kinase inhibition. ( A ) Serum withdrawal rapidly decreased hepcidin transcript levels in Huh7 cells. ( B ) After 40 h of FCS reduction from 10% to 0.4%, hepcidin transcripts were hardly detectable by RPA in HepG2 cells, whereas hypoxic IGFBP1 induction was not affected. Representative of three independent experiments. U6sn RNA served as loading control. ( C ) Exposure of Huh7 cells to protein kinase inhibitors revealed that the pan kinase inhibitor staurosporine (stauro; 0.5 µM) and the PI3 kinase inhibitor LY294002 (LY; 10 µM) reduced hepcidin expression similar to serum deprivation, whereas the p38 SAP kinase inhibitor SB202190 (SB; 10 µM) had no effect and the MEK1/2 inhibitor UO126 (1 µM) even increased HAMP/18S ratios. Data are results of qRT PCR analyses and given as means of three independent experiments±SEM; *p

    Journal: PLoS ONE

    Article Title: Evidence for a Lack of a Direct Transcriptional Suppression of the Iron Regulatory Peptide Hepcidin by Hypoxia-Inducible Factors

    doi: 10.1371/journal.pone.0007875

    Figure Lengend Snippet: Response of hepcidin transcript levels to serum deprivation and protein kinase inhibition. ( A ) Serum withdrawal rapidly decreased hepcidin transcript levels in Huh7 cells. ( B ) After 40 h of FCS reduction from 10% to 0.4%, hepcidin transcripts were hardly detectable by RPA in HepG2 cells, whereas hypoxic IGFBP1 induction was not affected. Representative of three independent experiments. U6sn RNA served as loading control. ( C ) Exposure of Huh7 cells to protein kinase inhibitors revealed that the pan kinase inhibitor staurosporine (stauro; 0.5 µM) and the PI3 kinase inhibitor LY294002 (LY; 10 µM) reduced hepcidin expression similar to serum deprivation, whereas the p38 SAP kinase inhibitor SB202190 (SB; 10 µM) had no effect and the MEK1/2 inhibitor UO126 (1 µM) even increased HAMP/18S ratios. Data are results of qRT PCR analyses and given as means of three independent experiments±SEM; *p

    Article Snippet: To identify kinase pathways that were driving basal hepcidin expression in cultured cells in the presence of FCS, the following inhibitors were used: the phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 (10 µM, Calbiochem), the p38 stress-activated protein kinase (SAPK) inhibitor SB202190 (10 µM, Sigma), the mitogen-activated protein kinase kinase (MEK)1/2 inhibitor UO126 (1 µM, Cell Signaling) and the pan kinase inhibitor staurosporine (0.5 µM, Sigma).

    Techniques: Inhibition, Recombinase Polymerase Amplification, Expressing, Quantitative RT-PCR

    SB431542 exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p

    Journal: Stem Cells (Dayton, Ohio)

    Article Title: Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial‐Like Cells

    doi: 10.1002/stem.2750

    Figure Lengend Snippet: SB431542 exposure during early differentiation enhanced the hiPSC differentiation to corneal epithelial progenitors. Schematic outline for optimization experiment using SB431542 (A) . Quantitative real‐time polymerase chain reaction analysis of the putative limbal stem cells ( ΔNp63 ) gene for SB‐Ad3 cells from G5 subgroups on day 20 of optimization experiment following different durations of SB431542 exposure (B) . Colony forming efficiency assays for the SB431542 exposed and unexposed G5 subgroups on day 20 (C) . Data presented as mean ± SEM, n = 3. *, statistically different compared with untreated group. **, p

    Article Snippet: SB431542 (10 µM) was added for 1, 2, and 3 days to the differentiation media in combination with BMP4, RA, and EGF as detailed in Figure A. qRT‐PCR analysis at day 20 indicated the highest expression of ΔNp63 in groups treated for 2 and 3 days with SB431542 (Fig. B).

    Techniques: Real-time Polymerase Chain Reaction

    Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), syntaxin (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain

    Journal: Biophysical Journal

    Article Title: Simulations Reveal Multiple Intermediates in the Unzipping Mechanism of Neuronal SNARE Complex

    doi: 10.1016/j.bpj.2018.08.043

    Figure Lengend Snippet: Schematic view of the SNARE complex. The four parallel helices of synaptobrevin (Sb), syntaxin (Sx), and SNAP25 form the core bundle of the complex. One arginine and three glutamine residues constitute the ionic layer, which divides the C-terminal domain

    Article Snippet: Synaptobrevin (Sb) and syntaxin (Sx) both contribute to this complex with one α -helix, whereas SNAP25 consists of two α -helical motifs connected by an unstructured linker loop.

    Techniques:

    Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on fibronectin-coated substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

    Journal: PLoS ONE

    Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

    doi: 10.1371/journal.pone.0052627

    Figure Lengend Snippet: Silencing of Exo70 leads to Cav1 accumulation in focal adhesions. Mock-treated ( A ) or Exo70-depleted cells ( B ) were detached for 1 h and replated on fibronectin-coated substrate for 3 h, and then fixed and stained for endogenous vinculin and Cav1. Scale bars, 5 µm. Co-localization between Cav1 and vinculin was quantified in a 20-pixel width region from the cell periphery and compared in Exo70-depleted with two independent siRNAs vs . mock-treated cells ( C ). Results in panel C are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

    Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

    Techniques: Staining

    Exo70 redistributes in Cav1-positive compartments upon cell detachment. ( A ) Hela cells expressing Cav1-mRFP and Exo70-GFP were kept in suspension for 1 h and replated on fibronectin for 3 h, and then visualized by confocal dual-colour spinning-disk microscopy (see corresponding movie S1). Arrow points to a dynamic Cav1- and Exo70-positive vesicle. Bottom panel shows selected frames from the time-lapse series corresponding to the boxed region in the upper panel. Time is given in second. ( B ) Hela cells expressing Cav1-GFP and cavin-1-mRFP were treated as in panel A. Inset shows higher magnification of region indicated by an arrow. Scale bars, 5 µm.

    Journal: PLoS ONE

    Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

    doi: 10.1371/journal.pone.0052627

    Figure Lengend Snippet: Exo70 redistributes in Cav1-positive compartments upon cell detachment. ( A ) Hela cells expressing Cav1-mRFP and Exo70-GFP were kept in suspension for 1 h and replated on fibronectin for 3 h, and then visualized by confocal dual-colour spinning-disk microscopy (see corresponding movie S1). Arrow points to a dynamic Cav1- and Exo70-positive vesicle. Bottom panel shows selected frames from the time-lapse series corresponding to the boxed region in the upper panel. Time is given in second. ( B ) Hela cells expressing Cav1-GFP and cavin-1-mRFP were treated as in panel A. Inset shows higher magnification of region indicated by an arrow. Scale bars, 5 µm.

    Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

    Techniques: Expressing, Microscopy

    Actin and microtubule cytoskeletons act at distinct steps of Cav1 trafficking. ( A, B ) Hela cells expressing Cav1-mRFP maintained in suspension for 1 h, were replated on fibronectin for 3 h in the presence of 10 µM nocodazole (panel A) or 10 µg/ml cytochalasin-B (panel B) for 30 min. Cells were then analyzed by time-lapse confocal spinning disk microscopy. The right panels represent selected frames from the time-lapse series (time is given in second). Arrows point to Cav1-positive intracellular vesicles. See corresponding movie S2 (panel A) and movie 3 (panel B). Scale bars, 5 µm.

    Journal: PLoS ONE

    Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

    doi: 10.1371/journal.pone.0052627

    Figure Lengend Snippet: Actin and microtubule cytoskeletons act at distinct steps of Cav1 trafficking. ( A, B ) Hela cells expressing Cav1-mRFP maintained in suspension for 1 h, were replated on fibronectin for 3 h in the presence of 10 µM nocodazole (panel A) or 10 µg/ml cytochalasin-B (panel B) for 30 min. Cells were then analyzed by time-lapse confocal spinning disk microscopy. The right panels represent selected frames from the time-lapse series (time is given in second). Arrows point to Cav1-positive intracellular vesicles. See corresponding movie S2 (panel A) and movie 3 (panel B). Scale bars, 5 µm.

    Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

    Techniques: Activated Clotting Time Assay, Expressing, Microscopy

    Exo70 is required for Cav1 transport to the plasma membrane. ( A, B ) Hela cells expressing Cav1-mRFP and α5-integrin-GFP either mock-treated (A) or treated with a specific siRNA to silencing Exo70 (B) were maintained in suspension for 1 h and replated on fibronectin for 3 h, and visualized using time-lapse spinning disk microscopy. Scale bars, 5 µm. ( A′, B′ ) Intensity profile of Cav1 (red) and α5-integrin (green) along the white lines shown in panel A and B. ( C ) Co-localization analysis of Cav1-mRFP and α5-integrin-GFP in cells as in panels A and B in a 20-pixel width region along the cell periphery of mock- or Exo70 siRNA-depleted cells using two specific siRNAs (siRNA 7c and 7d). Results are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

    Journal: PLoS ONE

    Article Title: Exo70 Subunit of the Exocyst Complex Is Involved in Adhesion-Dependent Trafficking of Caveolin-1

    doi: 10.1371/journal.pone.0052627

    Figure Lengend Snippet: Exo70 is required for Cav1 transport to the plasma membrane. ( A, B ) Hela cells expressing Cav1-mRFP and α5-integrin-GFP either mock-treated (A) or treated with a specific siRNA to silencing Exo70 (B) were maintained in suspension for 1 h and replated on fibronectin for 3 h, and visualized using time-lapse spinning disk microscopy. Scale bars, 5 µm. ( A′, B′ ) Intensity profile of Cav1 (red) and α5-integrin (green) along the white lines shown in panel A and B. ( C ) Co-localization analysis of Cav1-mRFP and α5-integrin-GFP in cells as in panels A and B in a 20-pixel width region along the cell periphery of mock- or Exo70 siRNA-depleted cells using two specific siRNAs (siRNA 7c and 7d). Results are the average of mean percentage of co-localization ± s.e.m. of three experiments. ** P

    Article Snippet: For spinning disk microscopy, HeLa cells plated onto a glass-bottom dish coated with fibronectin (Sigma, 10 µg/ml) and transfected with the indicated constructs.

    Techniques: Expressing, Microscopy

    Inactivation of Evl in unconfined MV D7 cells diminishes cell contractile energy. ( A ) Quantifications of contractile energy normalized by cellular area and measured for MV D7 , MVE-KO and Evl-rescue of MVE-KO cells plated on plain polyacrylamide micropatterns coated with fibronectin. ***p≤0.001 or n.s., not significant by Kruskal-Wallis test and Dunn’s Multiple Comparison test. n, number of cells analyzed. Boxes in box plots indicate 50% (25–75%) and whiskers (5–95%) of all measurements, with dashed black lines depicting the medians, arithmetic means are highlighted in red. ( B ) Images depicting representative traction force field representations from cells of distinct genotypes, as indicated. Force scale bar is in Pascal and arrows represent local force magnitude and orientation. Source data for details of contractile energies Figure 8—figure supplement 1 .

    Journal: eLife

    Article Title: Loss of Ena/VASP interferes with lamellipodium architecture, motility and integrin-dependent adhesion

    doi: 10.7554/eLife.55351

    Figure Lengend Snippet: Inactivation of Evl in unconfined MV D7 cells diminishes cell contractile energy. ( A ) Quantifications of contractile energy normalized by cellular area and measured for MV D7 , MVE-KO and Evl-rescue of MVE-KO cells plated on plain polyacrylamide micropatterns coated with fibronectin. ***p≤0.001 or n.s., not significant by Kruskal-Wallis test and Dunn’s Multiple Comparison test. n, number of cells analyzed. Boxes in box plots indicate 50% (25–75%) and whiskers (5–95%) of all measurements, with dashed black lines depicting the medians, arithmetic means are highlighted in red. ( B ) Images depicting representative traction force field representations from cells of distinct genotypes, as indicated. Force scale bar is in Pascal and arrows represent local force magnitude and orientation. Source data for details of contractile energies Figure 8—figure supplement 1 .

    Article Snippet: Cells were seeded onto 35 mm glass bottom dishes (Ibidi, Planegg-Martinsried, Germany) coated with either 25 µg/mL laminin (Sigma) in case of B16-F1 cells and derived clones, or with 10 µg/mL fibronectin (Roche, Penzberg, Germany) in case of MVD7 , NIH 3T3 and their derivatives, and maintained in imaging medium composed of F-12 Ham Nutrient Mixture with 25 mM HEPES (Sigma), the latter to compensate for the lack of CO2 , and supplemented with 10% FBS (Biowest, Nuaillé, France), 1% Penicillin-Streptomycin (Biowest), 2 mM stable L-glutamine (Biowest), and 2.7 g/L D-glucose (Carl Roth) in an Ibidi Heating System at 37°C.

    Techniques: