gfp Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore gfp
    Peripheral nervous system transduction. Representative sections of dorsal root ganglia at three levels (cervical, thoracic, lumbar) were stained to characterize transduced neuronal subtypes. <t>GFP-positive</t> cells were colocalized with sensory neuron (GFP
    Gfp, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp/product/Millipore
    Average 99 stars, based on 7375 article reviews
    Price from $9.99 to $1999.99
    gfp - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    Shanghai Genechem green fuorescent protein gfp gene
    Peripheral nervous system transduction. Representative sections of dorsal root ganglia at three levels (cervical, thoracic, lumbar) were stained to characterize transduced neuronal subtypes. <t>GFP-positive</t> cells were colocalized with sensory neuron (GFP
    Green Fuorescent Protein Gfp Gene, supplied by Shanghai Genechem, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/green fuorescent protein gfp gene/product/Shanghai Genechem
    Average 92 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    green fuorescent protein gfp gene - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    88
    The Jackson Laboratory green florescent protein gfp
    Detection of <t>GFP,</t> by fluorescence, in the lungs of adult mice. A shows a lung section from a saline-treated <t>transgenic</t> mouse. B and C show lung sections after PPE-induced injury, of a transgenic and wild-type mouse, respectively. Note the loss of tissue and disruption of normal lung architecture of PPE-treated lungs. There does not appear to be a change in the concentration of highly fluorescent tissue in the injured (emphysematous) areas. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.
    Green Florescent Protein Gfp, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/green florescent protein gfp/product/The Jackson Laboratory
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    green florescent protein gfp - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    88
    InvivoGen green florescent protein gfp
    Detection of <t>GFP,</t> by fluorescence, in the lungs of adult mice. A shows a lung section from a saline-treated <t>transgenic</t> mouse. B and C show lung sections after PPE-induced injury, of a transgenic and wild-type mouse, respectively. Note the loss of tissue and disruption of normal lung architecture of PPE-treated lungs. There does not appear to be a change in the concentration of highly fluorescent tissue in the injured (emphysematous) areas. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.
    Green Florescent Protein Gfp, supplied by InvivoGen, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/green florescent protein gfp/product/InvivoGen
    Average 88 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    green florescent protein gfp - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    93
    Thermo Fisher green fluorescent protein gfp
    Cell cycle exit increased by inhibition of <t>β-catenin/TCF</t> signaling. A , Animals were electroporated with expression plasmids for <t>GFP</t> alone or coelectroporated with DNTCF-4 or ICAT, and then exposed to a single pulse label of BrdU 24 h before killing.
    Green Fluorescent Protein Gfp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/green fluorescent protein gfp/product/Thermo Fisher
    Average 93 stars, based on 1194 article reviews
    Price from $9.99 to $1999.99
    green fluorescent protein gfp - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc gfp
    Subcellular localization of K3 is not different from wildtype <t>AID.</t> (A) Schematic representation of <t>AID-GFP</t> fusion constructs used for localization assays. GFP is fused to the C-terminus of AID or mutant K3. (B) HEK293T cells were transiently transfected with the respective GFP-fusion constructs. Two days post transfection, cells were treated as indicated with Leptomycin B (LMB) for 3 hours. Cells were fixed and localization of the respective GFP fusion proteins visualized using fluorescence microscopy. (C) The k46 B cell line stably expressing AID-GFP or K3-GFP fusion proteins were treated with or without Leptomycin B (LMB) for 3 hours and localization of GFP fusion constructs was determined using fluorescence microscopy.
    Gfp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1850 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1850 article reviews
    Price from $9.99 to $1999.99
    gfp - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    94
    R&D Systems gfp
    Cardiac mural cells originate from mesenchymal progenitors. ( a – g ) Clonal analysis using Pdgfrb(BAC)-CreERT2 Rosa26-mTmG mice. ( a ) Experimental strategy indicating the stages of 4-hydroxytamoxifen (4-OHT) administration and analysis. ( b ) <t>GFP+</t> cells (green, arrows) in AVC at 8 hours after 4-OHT administration. PDGFRβ (red) immunostaining and unrecombined cells (tdTomato, white/red) are shown. ( c , d ) Overview of GFP+ cell distribution in the indicated regions of E14.5 heart ( c ). At higher magnification, GFP+ PDGFRβ+ cells (arrows) in the myocardium were found in association with isolectin B4-labelled vessels (blue) ( d ). ( e ) Arrows indicate representative GFP+ cells, which were <t>PDGFRα+</t> (red) in the E10.5 AVC and E14.5 myocardium, but PDGFRα- and located at isolectin B4+ vessels (blue) in E16.5 myocardium. ( f ) Pdgfrb(BAC)-CreERT2 -labelled cell clones (GFP, green) gave rise to SM22α+ mural cells (red, arrow) at E16.5. ECs, isolectin B4 (blue). ( g ) Pdgfrb(BAC)-CreERT2 -marked cell clones (GFP, green) were identified as αSMA+ vSMCs (red, arrows) at E18.5. Arrowheads mark a αSMA- perivascular cells. ECs, isolectin B4 (blue).
    Gfp, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp/product/R&D Systems
    Average 94 stars, based on 182 article reviews
    Price from $9.99 to $1999.99
    gfp - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    88
    InvivoGen pmono hygro gfp pmhg
    Cardiac mural cells originate from mesenchymal progenitors. ( a – g ) Clonal analysis using Pdgfrb(BAC)-CreERT2 Rosa26-mTmG mice. ( a ) Experimental strategy indicating the stages of 4-hydroxytamoxifen (4-OHT) administration and analysis. ( b ) <t>GFP+</t> cells (green, arrows) in AVC at 8 hours after 4-OHT administration. PDGFRβ (red) immunostaining and unrecombined cells (tdTomato, white/red) are shown. ( c , d ) Overview of GFP+ cell distribution in the indicated regions of E14.5 heart ( c ). At higher magnification, GFP+ PDGFRβ+ cells (arrows) in the myocardium were found in association with isolectin B4-labelled vessels (blue) ( d ). ( e ) Arrows indicate representative GFP+ cells, which were <t>PDGFRα+</t> (red) in the E10.5 AVC and E14.5 myocardium, but PDGFRα- and located at isolectin B4+ vessels (blue) in E16.5 myocardium. ( f ) Pdgfrb(BAC)-CreERT2 -labelled cell clones (GFP, green) gave rise to SM22α+ mural cells (red, arrow) at E16.5. ECs, isolectin B4 (blue). ( g ) Pdgfrb(BAC)-CreERT2 -marked cell clones (GFP, green) were identified as αSMA+ vSMCs (red, arrows) at E18.5. Arrowheads mark a αSMA- perivascular cells. ECs, isolectin B4 (blue).
    Pmono Hygro Gfp Pmhg, supplied by InvivoGen, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmono hygro gfp pmhg/product/InvivoGen
    Average 88 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    pmono hygro gfp pmhg - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc gfp 2555
    The amino acid dependence of mTOR complex 1 binding to recombinant Rag heterodimers in HeLa cells stably expressing <t>GFP-streptag-RagC</t> variants at differing abundances. HeLa cells stably expressing GFP-streptag, GFP-streptag-RagC WT , or GFP-streptag-RagC S75L were selected by cell sorting for GFP abundance, and the expression of the full-length recombinant protein was estimated by GFP immunoblot analysis of lysates. The cells were incubated in fresh DMEM for 1 h and then deprived of amino acids by incubation in DPBS for 1.5 h. The medium of plates AA- was replaced with fresh DPBS, whereas that of plates AA+ was replaced with DMEM, with harvest 10 min thereafter. Strep-Tactin pull-downs ( PD ) and the lysates were analyzed by immunoblotting for endogenous ( Endo ) Raptor and RagA, for GFP, and for <t>4E-BP(T37P/T46P).</t>
    Gfp 2555, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp 2555/product/Cell Signaling Technology Inc
    Average 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    gfp 2555 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    88
    Vector Biolabs gfp ad gfp
    Ectopic expression of <t>HNF6</t> in acinar cells is associated with induction of ductal genes and repression of acinar genes in vitro . (A) Relative expression of ductal (left) and acinar (right) genes in cultured 266-6 cells. Cells were transduced with an adenovirus expressing <t>GFP</t> (Ad-GFP), HNF6 (Ad-HNF6), or Sox9 (Ad-Sox9) and gene expression was analyzed by Q-RT-PCR (data are means +/− SEM; n= 4; *, p
    Gfp Ad Gfp, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp ad gfp/product/Vector Biolabs
    Average 88 stars, based on 47 article reviews
    Price from $9.99 to $1999.99
    gfp ad gfp - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    90
    Aldevron gfp gwiz gfp
    Ectopic expression of <t>HNF6</t> in acinar cells is associated with induction of ductal genes and repression of acinar genes in vitro . (A) Relative expression of ductal (left) and acinar (right) genes in cultured 266-6 cells. Cells were transduced with an adenovirus expressing <t>GFP</t> (Ad-GFP), HNF6 (Ad-HNF6), or Sox9 (Ad-Sox9) and gene expression was analyzed by Q-RT-PCR (data are means +/− SEM; n= 4; *, p
    Gfp Gwiz Gfp, supplied by Aldevron, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp gwiz gfp/product/Aldevron
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    gfp gwiz gfp - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    93
    InvivoGen pmono neo gfp vector
    Ectopic expression of <t>HNF6</t> in acinar cells is associated with induction of ductal genes and repression of acinar genes in vitro . (A) Relative expression of ductal (left) and acinar (right) genes in cultured 266-6 cells. Cells were transduced with an adenovirus expressing <t>GFP</t> (Ad-GFP), HNF6 (Ad-HNF6), or Sox9 (Ad-Sox9) and gene expression was analyzed by Q-RT-PCR (data are means +/− SEM; n= 4; *, p
    Pmono Neo Gfp Vector, supplied by InvivoGen, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmono neo gfp vector/product/InvivoGen
    Average 93 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    pmono neo gfp vector - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    85
    Becton Dickinson gfp gfp bd
    <t>GFP-BD</t> interaction with acetylated <t>mCherry-H4</t> in the nucleus of HEK293 live cells using a single-photon counting method. ( A ) Steady-state intensity image of GFP-BD expressed alone in HEK293 cell ( green ). The corresponding GFP-BD fluorescence decay ( green
    Gfp Gfp Bd, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp gfp bd/product/Becton Dickinson
    Average 85 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    gfp gfp bd - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

    93
    System Biosciences Inc gfp mito gfp
    Mitochondria are transferred from iPSC-MSCs to injured PC12 cells under CoCl 2 challenge. (A) TNTs formed between PC12 cells (in green) and iPSC-MSCs. Scale bar: 10 μm. (B) The number of iPSC-MSC TNTs was greatly enhanced under CoCl 2 challenge. (C) Mitochondria were transferred from <t>Mito-GFP-iPSC-MSCs</t> (in green) to Celltrace-labelled-PC12 cells (in blue) via TNTs. Scale bar: 10 μm. (D) The efficiency of mitochondrial transfer from iPSC-MSCs to PC12 cells was dramatically enhanced under CoCl 2 treatment compared with no CoCl 2 treatment. (E) ATP levels were dramatically increased in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. (F) Apoptosis was dramatically reduced in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. Data are expressed as the mean ± SEM ( n = 3; unpaired Student’s t -test). All experiments were conducted in triplicate. * P
    Gfp Mito Gfp, supplied by System Biosciences Inc, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp mito gfp/product/System Biosciences Inc
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    gfp mito gfp - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    88
    Thermo Fisher a gfp
    Immunofluorescent stainings against <t>GABA</t> . GABA immunoreactivity shown alone in gray (A2,B2,C2,D2) , or (in magenta) overlay with Ccap-Gal4-driven expression of <t>GFP</t> (green, A1,B1,C1,D1 ). In the brain [ (A1,A2) , projection of 8 confocal slices containing the IN brain ] and the sog [ (B1,B2) , projection of 2 confocal slices containing the IN sog ], all N CCAP are GABA-immunonegative. In some but not all preparations of the thoracic and abdominal neuromeres, GABA immunoreactivity was found in one N CCAP per hemineuromere. (C1,C2) While the N CCAP on the right side of a4 and a5 are GABA-immunonegative (arrowheads), there is one N CCAP on the left side in a5 that shows weak but distinct GABA labeling (arrow, projection of 2 confocal slices). (D1,D2) shows a projection of 8 confocal slices of another preparation, where the N CCAP in a1, a3 and the anterior N CCAP in a4 appear to be GABA-immunonegative (upper arrows). One of the N CCAP in a4 (lowermost arrow) however shows weak GABA immunoreactivity. Scale bars = 50 μm.
    A Gfp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a gfp/product/Thermo Fisher
    Average 88 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    a gfp - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    88
    KeraFAST gfps
    Immunofluorescent stainings against <t>GABA</t> . GABA immunoreactivity shown alone in gray (A2,B2,C2,D2) , or (in magenta) overlay with Ccap-Gal4-driven expression of <t>GFP</t> (green, A1,B1,C1,D1 ). In the brain [ (A1,A2) , projection of 8 confocal slices containing the IN brain ] and the sog [ (B1,B2) , projection of 2 confocal slices containing the IN sog ], all N CCAP are GABA-immunonegative. In some but not all preparations of the thoracic and abdominal neuromeres, GABA immunoreactivity was found in one N CCAP per hemineuromere. (C1,C2) While the N CCAP on the right side of a4 and a5 are GABA-immunonegative (arrowheads), there is one N CCAP on the left side in a5 that shows weak but distinct GABA labeling (arrow, projection of 2 confocal slices). (D1,D2) shows a projection of 8 confocal slices of another preparation, where the N CCAP in a1, a3 and the anterior N CCAP in a4 appear to be GABA-immunonegative (upper arrows). One of the N CCAP in a4 (lowermost arrow) however shows weak GABA immunoreactivity. Scale bars = 50 μm.
    Gfps, supplied by KeraFAST, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfps/product/KeraFAST
    Average 88 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    gfps - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    93
    R&D Systems gfp antibody
    Immunofluorescent stainings against <t>GABA</t> . GABA immunoreactivity shown alone in gray (A2,B2,C2,D2) , or (in magenta) overlay with Ccap-Gal4-driven expression of <t>GFP</t> (green, A1,B1,C1,D1 ). In the brain [ (A1,A2) , projection of 8 confocal slices containing the IN brain ] and the sog [ (B1,B2) , projection of 2 confocal slices containing the IN sog ], all N CCAP are GABA-immunonegative. In some but not all preparations of the thoracic and abdominal neuromeres, GABA immunoreactivity was found in one N CCAP per hemineuromere. (C1,C2) While the N CCAP on the right side of a4 and a5 are GABA-immunonegative (arrowheads), there is one N CCAP on the left side in a5 that shows weak but distinct GABA labeling (arrow, projection of 2 confocal slices). (D1,D2) shows a projection of 8 confocal slices of another preparation, where the N CCAP in a1, a3 and the anterior N CCAP in a4 appear to be GABA-immunonegative (upper arrows). One of the N CCAP in a4 (lowermost arrow) however shows weak GABA immunoreactivity. Scale bars = 50 μm.
    Gfp Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp antibody/product/R&D Systems
    Average 93 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    gfp antibody - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    gfp  (Abcam)
    93
    Abcam gfp
    BAY 11-7082 suppresses the LPS- or IL-1-stimulated formation of <t>K63-pUb</t> chains and the DNA damage response ( A and B ) The experiment was carried out as in Figure 2 , except that the K63-pUb chains formed in response to LPS ( A ) or IL-1β ( B ) were captured on Halo-NEMO from 6 mg (RAW cells) or 3 mg (IL-1R cells) of cell extract protein as described in the Experimental section. The K63-pUb chains were identified by immunoblotting with a specific antibody. Further aliquots of the cell extract were immunoblotted for IKKβ phosphorylation, p105 phosphorylation and GAPDH as in Figure 2 . ( C ) Indirect immunofluorescence images of U2OS cells transiently expressing <t>GFP–RAP80[1–200].</t> Cells were incubated for 1 h with or without BAY 11-7082 (15 μM) and either exposed to ionizing radiation (IR) or not exposed. GFP–RAP80 or γH2AX were visualized using anti-GFP and anti-γH2AX antibodies respectively, and nuclei were stained with DAPI.
    Gfp, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 8593 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp/product/Abcam
    Average 93 stars, based on 8593 article reviews
    Price from $9.99 to $1999.99
    gfp - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    a gfp  (Abcam)
    88
    Abcam a gfp
    Effect of AAV2.5-driven <t>SERCA2a</t> gene transfer on post-injury vascula healing Two groups of animals were analyzed: sham-operated control, injured and <t>AAV2.5-GFP</t> (n=10) or AAV2.5-SERCA2a (n=8) infected carotid artery 1 month after surgery. A . Representative hematoxylin/eosin staining of carotid artery cross-section. Objective X10 (upper panel), X60 (lower panel); ni – neointima; m – media; a – adventitia, lm -lumen. B . Morphometric analysis of carotid artery cross-sections. Bars represent the mean ± SEM of mean values obtained for each animal. At least 5 individual measures were performed for each animal on different carotid cross sections.
    A Gfp, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a gfp/product/Abcam
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    a gfp - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    Image Search Results


    Peripheral nervous system transduction. Representative sections of dorsal root ganglia at three levels (cervical, thoracic, lumbar) were stained to characterize transduced neuronal subtypes. GFP-positive cells were colocalized with sensory neuron (GFP

    Journal: Human Gene Therapy

    Article Title: Strong Cortical and Spinal Cord Transduction After AAV7 and AAV9 Delivery into the Cerebrospinal Fluid of Nonhuman Primates

    doi: 10.1089/hum.2013.005

    Figure Lengend Snippet: Peripheral nervous system transduction. Representative sections of dorsal root ganglia at three levels (cervical, thoracic, lumbar) were stained to characterize transduced neuronal subtypes. GFP-positive cells were colocalized with sensory neuron (GFP

    Article Snippet: Additionally, to determine the type of sensory DRG cell bodies transduced, DRG sections were costained for GFP and either β-III tubulin (mouse monoclonal, clone TUJ1, 1:100, MAB1637 Millipore) or NF-200 (mouse monoclonal, clone RT-97, 1:100, CBL212 Millipore) to identify myelinated fibers and neurons.

    Techniques: Transduction, Staining

    Cellular phenotypic analysis of AAV7-GFP transduced cells. Brain sections were costained against GFP and neuronal (NeuN) or astrocytic (GFAP, S100) markers. Both neurons (a) as well as fibrous (b) and protoplasmic astrocytes (c) were transduced. Arrows

    Journal: Human Gene Therapy

    Article Title: Strong Cortical and Spinal Cord Transduction After AAV7 and AAV9 Delivery into the Cerebrospinal Fluid of Nonhuman Primates

    doi: 10.1089/hum.2013.005

    Figure Lengend Snippet: Cellular phenotypic analysis of AAV7-GFP transduced cells. Brain sections were costained against GFP and neuronal (NeuN) or astrocytic (GFAP, S100) markers. Both neurons (a) as well as fibrous (b) and protoplasmic astrocytes (c) were transduced. Arrows

    Article Snippet: Additionally, to determine the type of sensory DRG cell bodies transduced, DRG sections were costained for GFP and either β-III tubulin (mouse monoclonal, clone TUJ1, 1:100, MAB1637 Millipore) or NF-200 (mouse monoclonal, clone RT-97, 1:100, CBL212 Millipore) to identify myelinated fibers and neurons.

    Techniques:

    Glia-like cells transduced in dorsal root ganglia. Double fluorescent immunohistochemistry revealed some GFP-positive satellite glia cells proximal to β-III tubulin–positive DRG neurons. Scale bar: 50 μm.

    Journal: Human Gene Therapy

    Article Title: Strong Cortical and Spinal Cord Transduction After AAV7 and AAV9 Delivery into the Cerebrospinal Fluid of Nonhuman Primates

    doi: 10.1089/hum.2013.005

    Figure Lengend Snippet: Glia-like cells transduced in dorsal root ganglia. Double fluorescent immunohistochemistry revealed some GFP-positive satellite glia cells proximal to β-III tubulin–positive DRG neurons. Scale bar: 50 μm.

    Article Snippet: Additionally, to determine the type of sensory DRG cell bodies transduced, DRG sections were costained for GFP and either β-III tubulin (mouse monoclonal, clone TUJ1, 1:100, MAB1637 Millipore) or NF-200 (mouse monoclonal, clone RT-97, 1:100, CBL212 Millipore) to identify myelinated fibers and neurons.

    Techniques: Immunohistochemistry

    Detection of GFP, by fluorescence, in the lungs of adult mice. A shows a lung section from a saline-treated transgenic mouse. B and C show lung sections after PPE-induced injury, of a transgenic and wild-type mouse, respectively. Note the loss of tissue and disruption of normal lung architecture of PPE-treated lungs. There does not appear to be a change in the concentration of highly fluorescent tissue in the injured (emphysematous) areas. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Engraftment of Neonatal Lung Fibroblasts into the Normal and Elastase-Injured Lung

    doi: 10.1165/rcmb.2004-0319OC

    Figure Lengend Snippet: Detection of GFP, by fluorescence, in the lungs of adult mice. A shows a lung section from a saline-treated transgenic mouse. B and C show lung sections after PPE-induced injury, of a transgenic and wild-type mouse, respectively. Note the loss of tissue and disruption of normal lung architecture of PPE-treated lungs. There does not appear to be a change in the concentration of highly fluorescent tissue in the injured (emphysematous) areas. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.

    Article Snippet: Neonatal lung fibroblasts also referred to as lung interstitial cells (LIC) were isolated from the lungs of 10-d-old C57BL/6J and green florescent protein (GFP)-expressing transgenic mice (C57BL/6(ACTbEGFP)1Osb/J; Jackson Laboratory, Bar Harbor, ME) by digestion with 0.25% typsin IX and collagenase type II (Sigma-Aldrich, St. Louis, MO) as previously described ( , ).

    Techniques: Fluorescence, Mouse Assay, Transgenic Assay, Concentration Assay

    Detection of GFP in 10-d-old mouse pups. ( A ) Unstained lung tissue of a wild-type pup. B demonstrates the green fluorescence of the unstained lung of a GFP-transgenic pup that is heterozygous for the GFP gene under the same illumination. It can be seen that most cells of the transgenic lung display the green fluorescence of the GFP. The most notable exceptions are the red blood cells, seen in B as pink-colored cells within blood vessels. The cells with the greatest fluorescence are in the parenchyma, at the location of type II cells, and in the airway epithelium. C demonstrates immunostaining for GFP in a GFP-transgenic lung and detects the same distribution pattern for GFP, as with fluorescence illumination of unstained sections. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Engraftment of Neonatal Lung Fibroblasts into the Normal and Elastase-Injured Lung

    doi: 10.1165/rcmb.2004-0319OC

    Figure Lengend Snippet: Detection of GFP in 10-d-old mouse pups. ( A ) Unstained lung tissue of a wild-type pup. B demonstrates the green fluorescence of the unstained lung of a GFP-transgenic pup that is heterozygous for the GFP gene under the same illumination. It can be seen that most cells of the transgenic lung display the green fluorescence of the GFP. The most notable exceptions are the red blood cells, seen in B as pink-colored cells within blood vessels. The cells with the greatest fluorescence are in the parenchyma, at the location of type II cells, and in the airway epithelium. C demonstrates immunostaining for GFP in a GFP-transgenic lung and detects the same distribution pattern for GFP, as with fluorescence illumination of unstained sections. All panels are the same magnification; short (vertical) dimensions of panels are 436 μm.

    Article Snippet: Neonatal lung fibroblasts also referred to as lung interstitial cells (LIC) were isolated from the lungs of 10-d-old C57BL/6J and green florescent protein (GFP)-expressing transgenic mice (C57BL/6(ACTbEGFP)1Osb/J; Jackson Laboratory, Bar Harbor, ME) by digestion with 0.25% typsin IX and collagenase type II (Sigma-Aldrich, St. Louis, MO) as previously described ( , ).

    Techniques: Fluorescence, Transgenic Assay, Immunostaining

    Cell cycle exit increased by inhibition of β-catenin/TCF signaling. A , Animals were electroporated with expression plasmids for GFP alone or coelectroporated with DNTCF-4 or ICAT, and then exposed to a single pulse label of BrdU 24 h before killing.

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Cell-Autonomous ?-Catenin Signaling Regulates Cortical Precursor Proliferation

    doi: 10.1523/JNEUROSCI.3180-06.2006

    Figure Lengend Snippet: Cell cycle exit increased by inhibition of β-catenin/TCF signaling. A , Animals were electroporated with expression plasmids for GFP alone or coelectroporated with DNTCF-4 or ICAT, and then exposed to a single pulse label of BrdU 24 h before killing.

    Article Snippet: Cells were incubated at 37°C for 2 h, and then fixed with 4% paraformaldehyde for 10 min. Coverslips were then stained for β-catenin (BD Transduction Laboratories, Lexington, KY; mouse monoclonal), green fluorescent protein (GFP) (Invitrogen, Eugene, OR; rabbit polyclonal), and DNA (Hoechst 33342).

    Techniques: Inhibition, Expressing

    β-Catenin signaling undergoes dynamic changes during neuronal differentiation. E13.5 ventricularzone precursors coelectroporated with equal amounts of pCAG-mRFP or pTOPdGFP were analyzed after different intervals. At 24 h, GFP expression, representing

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Cell-Autonomous ?-Catenin Signaling Regulates Cortical Precursor Proliferation

    doi: 10.1523/JNEUROSCI.3180-06.2006

    Figure Lengend Snippet: β-Catenin signaling undergoes dynamic changes during neuronal differentiation. E13.5 ventricularzone precursors coelectroporated with equal amounts of pCAG-mRFP or pTOPdGFP were analyzed after different intervals. At 24 h, GFP expression, representing

    Article Snippet: Cells were incubated at 37°C for 2 h, and then fixed with 4% paraformaldehyde for 10 min. Coverslips were then stained for β-catenin (BD Transduction Laboratories, Lexington, KY; mouse monoclonal), green fluorescent protein (GFP) (Invitrogen, Eugene, OR; rabbit polyclonal), and DNA (Hoechst 33342).

    Techniques: Expressing

    β-Catenin protein is reduced after Cre-electroporation in floxed β-catenin mouse cortex. Floxed β-catenin cortical cells electroporated with Cre-Ires2-GFP have decreased immunoreactivity for β-catenin after 24 h ( A ) and

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Cell-Autonomous ?-Catenin Signaling Regulates Cortical Precursor Proliferation

    doi: 10.1523/JNEUROSCI.3180-06.2006

    Figure Lengend Snippet: β-Catenin protein is reduced after Cre-electroporation in floxed β-catenin mouse cortex. Floxed β-catenin cortical cells electroporated with Cre-Ires2-GFP have decreased immunoreactivity for β-catenin after 24 h ( A ) and

    Article Snippet: Cells were incubated at 37°C for 2 h, and then fixed with 4% paraformaldehyde for 10 min. Coverslips were then stained for β-catenin (BD Transduction Laboratories, Lexington, KY; mouse monoclonal), green fluorescent protein (GFP) (Invitrogen, Eugene, OR; rabbit polyclonal), and DNA (Hoechst 33342).

    Techniques: Electroporation

    Inhibition of β-catenin signaling causes premature exit from the ventricular zone. E13.5 brains coelectroporated with pCAG-GFP-DN-TCF4 and pCAG-mRFP ( A ) or pCAG-ICAT-IRES-EGFP and pCAG-mRFP ( B ) and analyzed at E16.5. A greater fraction of cells

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Cell-Autonomous ?-Catenin Signaling Regulates Cortical Precursor Proliferation

    doi: 10.1523/JNEUROSCI.3180-06.2006

    Figure Lengend Snippet: Inhibition of β-catenin signaling causes premature exit from the ventricular zone. E13.5 brains coelectroporated with pCAG-GFP-DN-TCF4 and pCAG-mRFP ( A ) or pCAG-ICAT-IRES-EGFP and pCAG-mRFP ( B ) and analyzed at E16.5. A greater fraction of cells

    Article Snippet: Cells were incubated at 37°C for 2 h, and then fixed with 4% paraformaldehyde for 10 min. Coverslips were then stained for β-catenin (BD Transduction Laboratories, Lexington, KY; mouse monoclonal), green fluorescent protein (GFP) (Invitrogen, Eugene, OR; rabbit polyclonal), and DNA (Hoechst 33342).

    Techniques: Inhibition

    β-Catenin signaling in cortical neural precursors. β-Catenin signaling in cortical ventricular zone revealed by expression of destabilized GFP controlled by TOP reporter elements. E13.5 precursors were electroporated with pTOP-dGFP and

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Cell-Autonomous ?-Catenin Signaling Regulates Cortical Precursor Proliferation

    doi: 10.1523/JNEUROSCI.3180-06.2006

    Figure Lengend Snippet: β-Catenin signaling in cortical neural precursors. β-Catenin signaling in cortical ventricular zone revealed by expression of destabilized GFP controlled by TOP reporter elements. E13.5 precursors were electroporated with pTOP-dGFP and

    Article Snippet: Cells were incubated at 37°C for 2 h, and then fixed with 4% paraformaldehyde for 10 min. Coverslips were then stained for β-catenin (BD Transduction Laboratories, Lexington, KY; mouse monoclonal), green fluorescent protein (GFP) (Invitrogen, Eugene, OR; rabbit polyclonal), and DNA (Hoechst 33342).

    Techniques: Expressing

    Subcellular localization of K3 is not different from wildtype AID. (A) Schematic representation of AID-GFP fusion constructs used for localization assays. GFP is fused to the C-terminus of AID or mutant K3. (B) HEK293T cells were transiently transfected with the respective GFP-fusion constructs. Two days post transfection, cells were treated as indicated with Leptomycin B (LMB) for 3 hours. Cells were fixed and localization of the respective GFP fusion proteins visualized using fluorescence microscopy. (C) The k46 B cell line stably expressing AID-GFP or K3-GFP fusion proteins were treated with or without Leptomycin B (LMB) for 3 hours and localization of GFP fusion constructs was determined using fluorescence microscopy.

    Journal: PLoS ONE

    Article Title: Lysine Residue at Position 22 of the AID Protein Regulates Its Class Switch Activity

    doi: 10.1371/journal.pone.0030667

    Figure Lengend Snippet: Subcellular localization of K3 is not different from wildtype AID. (A) Schematic representation of AID-GFP fusion constructs used for localization assays. GFP is fused to the C-terminus of AID or mutant K3. (B) HEK293T cells were transiently transfected with the respective GFP-fusion constructs. Two days post transfection, cells were treated as indicated with Leptomycin B (LMB) for 3 hours. Cells were fixed and localization of the respective GFP fusion proteins visualized using fluorescence microscopy. (C) The k46 B cell line stably expressing AID-GFP or K3-GFP fusion proteins were treated with or without Leptomycin B (LMB) for 3 hours and localization of GFP fusion constructs was determined using fluorescence microscopy.

    Article Snippet: Samples were blotted onto PVDF membranes (Millipore) and detected by Western blot analysis using polyclonal rabbit serum specific for human AID (Abcam), GFP (Cell Signaling), or actin (Cell Signaling) and HRP-conjugated secondary antibodies specific for rabbit IgG (Dako).

    Techniques: Construct, Mutagenesis, Transfection, Fluorescence, Microscopy, Stable Transfection, Expressing

    Somatic Hypermutation of IgV genes by wt AID and mutant K3. (A) Somatic hypermutation reflected as loss of IgM expression in AID −/− ΨV −/− IgM + DT40 cells. AID −/− ΨV −/− IgM + DT40 cells were stably transfected with plasmids encoding AID or K3 coupled to IRES-GFP expression. The percentage of sIgM-loss variants of GFP + cells is expressed as the median ± SE of multiple (n) independent clonal transfectants determined three weeks after transfection (untransfected control, n = 12; AID, n = 36; K3, n = 44). (B) Proportion of sequences carrying mutations in IgV genes of DT40 cells. IgV regions were cloned from cDNA of AID and K3 expressing DT40 transfectants. Segment sizes in the pie charts are proportional to the number of sequences carrying the number of mutations indicated in the periphery of each chart. The frequency of mutations per bp sequenced and the total number of independent sequences analyzed is indicated underneath and in the center of each chart, respectively. (C) The abundance of AID and K3 in DT40 transfectants was determined by Western blotting from cell extracts. Blots were reprobed for GFP expression and actin levels as loading control. (D) Proportion of sequences carrying Bcl6 mutations in AID deficient B cells retrovirally transfected with AID or K3 constructs, respectively. Bcl6 was cloned from genomic DNA of sorted mouse B cells, retrovirally transfected to express AID or K3, respectively. Pie charts were generated as described in (B).

    Journal: PLoS ONE

    Article Title: Lysine Residue at Position 22 of the AID Protein Regulates Its Class Switch Activity

    doi: 10.1371/journal.pone.0030667

    Figure Lengend Snippet: Somatic Hypermutation of IgV genes by wt AID and mutant K3. (A) Somatic hypermutation reflected as loss of IgM expression in AID −/− ΨV −/− IgM + DT40 cells. AID −/− ΨV −/− IgM + DT40 cells were stably transfected with plasmids encoding AID or K3 coupled to IRES-GFP expression. The percentage of sIgM-loss variants of GFP + cells is expressed as the median ± SE of multiple (n) independent clonal transfectants determined three weeks after transfection (untransfected control, n = 12; AID, n = 36; K3, n = 44). (B) Proportion of sequences carrying mutations in IgV genes of DT40 cells. IgV regions were cloned from cDNA of AID and K3 expressing DT40 transfectants. Segment sizes in the pie charts are proportional to the number of sequences carrying the number of mutations indicated in the periphery of each chart. The frequency of mutations per bp sequenced and the total number of independent sequences analyzed is indicated underneath and in the center of each chart, respectively. (C) The abundance of AID and K3 in DT40 transfectants was determined by Western blotting from cell extracts. Blots were reprobed for GFP expression and actin levels as loading control. (D) Proportion of sequences carrying Bcl6 mutations in AID deficient B cells retrovirally transfected with AID or K3 constructs, respectively. Bcl6 was cloned from genomic DNA of sorted mouse B cells, retrovirally transfected to express AID or K3, respectively. Pie charts were generated as described in (B).

    Article Snippet: Samples were blotted onto PVDF membranes (Millipore) and detected by Western blot analysis using polyclonal rabbit serum specific for human AID (Abcam), GFP (Cell Signaling), or actin (Cell Signaling) and HRP-conjugated secondary antibodies specific for rabbit IgG (Dako).

    Techniques: Mutagenesis, Expressing, Stable Transfection, Transfection, Clone Assay, Western Blot, Construct, Generated

    Effect of lysine to arginine mutations in the AID protein on CSR activity. (A) Schematic representation of AID highlighting the position of the lysine (K) residues. For CSR assays, untagged constructs were expressed bicistronically together with GFP separated by an internal ribosome entry site (IRES). (B) Representative FACS profile for CSR activity of individual Lys to Arg mutants. B cells from AID deficient mice were retrovirally transfected with the respective lysine AID mutant (Mx: empty vector) and CSR to IgG1 was determined upon stimulation of cells with LPS and IL-4. GFP expression of infected cells was driven from an IRES element located 3′ of the stop codon of the cloned AID mutant. The percentage in the upper right quadrants indicate the percentage of IgG1 + cells among the GFP + (i.e. retrovirally infected) population. (C) Results from three independent experiments of a screen of lysine mutants showing the percentage of IgG1 + cells among the GFP + population. (D) Results from 11 independent experiments comparing the CSR activity of AID and K3. (E) Western Blot on lysates from B cells retrovirally transfected with the respective constructs. Black arrowheads indicate specific band. White arrowheads indicate nonspecific band. (IB: immune blot).

    Journal: PLoS ONE

    Article Title: Lysine Residue at Position 22 of the AID Protein Regulates Its Class Switch Activity

    doi: 10.1371/journal.pone.0030667

    Figure Lengend Snippet: Effect of lysine to arginine mutations in the AID protein on CSR activity. (A) Schematic representation of AID highlighting the position of the lysine (K) residues. For CSR assays, untagged constructs were expressed bicistronically together with GFP separated by an internal ribosome entry site (IRES). (B) Representative FACS profile for CSR activity of individual Lys to Arg mutants. B cells from AID deficient mice were retrovirally transfected with the respective lysine AID mutant (Mx: empty vector) and CSR to IgG1 was determined upon stimulation of cells with LPS and IL-4. GFP expression of infected cells was driven from an IRES element located 3′ of the stop codon of the cloned AID mutant. The percentage in the upper right quadrants indicate the percentage of IgG1 + cells among the GFP + (i.e. retrovirally infected) population. (C) Results from three independent experiments of a screen of lysine mutants showing the percentage of IgG1 + cells among the GFP + population. (D) Results from 11 independent experiments comparing the CSR activity of AID and K3. (E) Western Blot on lysates from B cells retrovirally transfected with the respective constructs. Black arrowheads indicate specific band. White arrowheads indicate nonspecific band. (IB: immune blot).

    Article Snippet: Samples were blotted onto PVDF membranes (Millipore) and detected by Western blot analysis using polyclonal rabbit serum specific for human AID (Abcam), GFP (Cell Signaling), or actin (Cell Signaling) and HRP-conjugated secondary antibodies specific for rabbit IgG (Dako).

    Techniques: Activity Assay, Construct, FACS, Mouse Assay, Transfection, Mutagenesis, Plasmid Preparation, Expressing, Infection, Clone Assay, Western Blot

    Cardiac mural cells originate from mesenchymal progenitors. ( a – g ) Clonal analysis using Pdgfrb(BAC)-CreERT2 Rosa26-mTmG mice. ( a ) Experimental strategy indicating the stages of 4-hydroxytamoxifen (4-OHT) administration and analysis. ( b ) GFP+ cells (green, arrows) in AVC at 8 hours after 4-OHT administration. PDGFRβ (red) immunostaining and unrecombined cells (tdTomato, white/red) are shown. ( c , d ) Overview of GFP+ cell distribution in the indicated regions of E14.5 heart ( c ). At higher magnification, GFP+ PDGFRβ+ cells (arrows) in the myocardium were found in association with isolectin B4-labelled vessels (blue) ( d ). ( e ) Arrows indicate representative GFP+ cells, which were PDGFRα+ (red) in the E10.5 AVC and E14.5 myocardium, but PDGFRα- and located at isolectin B4+ vessels (blue) in E16.5 myocardium. ( f ) Pdgfrb(BAC)-CreERT2 -labelled cell clones (GFP, green) gave rise to SM22α+ mural cells (red, arrow) at E16.5. ECs, isolectin B4 (blue). ( g ) Pdgfrb(BAC)-CreERT2 -marked cell clones (GFP, green) were identified as αSMA+ vSMCs (red, arrows) at E18.5. Arrowheads mark a αSMA- perivascular cells. ECs, isolectin B4 (blue).

    Journal: Nature Communications

    Article Title: Endothelial cells are progenitors of cardiac pericytes and vascular smooth muscle cells

    doi: 10.1038/ncomms12422

    Figure Lengend Snippet: Cardiac mural cells originate from mesenchymal progenitors. ( a – g ) Clonal analysis using Pdgfrb(BAC)-CreERT2 Rosa26-mTmG mice. ( a ) Experimental strategy indicating the stages of 4-hydroxytamoxifen (4-OHT) administration and analysis. ( b ) GFP+ cells (green, arrows) in AVC at 8 hours after 4-OHT administration. PDGFRβ (red) immunostaining and unrecombined cells (tdTomato, white/red) are shown. ( c , d ) Overview of GFP+ cell distribution in the indicated regions of E14.5 heart ( c ). At higher magnification, GFP+ PDGFRβ+ cells (arrows) in the myocardium were found in association with isolectin B4-labelled vessels (blue) ( d ). ( e ) Arrows indicate representative GFP+ cells, which were PDGFRα+ (red) in the E10.5 AVC and E14.5 myocardium, but PDGFRα- and located at isolectin B4+ vessels (blue) in E16.5 myocardium. ( f ) Pdgfrb(BAC)-CreERT2 -labelled cell clones (GFP, green) gave rise to SM22α+ mural cells (red, arrow) at E16.5. ECs, isolectin B4 (blue). ( g ) Pdgfrb(BAC)-CreERT2 -marked cell clones (GFP, green) were identified as αSMA+ vSMCs (red, arrows) at E18.5. Arrowheads mark a αSMA- perivascular cells. ECs, isolectin B4 (blue).

    Article Snippet: The following primary antibodies were used: PECAM1 (553370, Pharmingen, 1:100), PDGFRβ (14-1402-82, eBioscience, 1:100), PDGFRα (14-1401-81, eBioscience, 1:100), isolectin B4 (B-1205, Vector, 1:100), NG2 (AB5320, Millipore, 1:100), PDGFRα (3164, Cell Signaling, 1:100), GFP (A21311, Invitrogen, 1:100), GFP (GFP-1010, Aves, 1:200), VE-cadherin (AF1002, R & D Systems, 1:100), CD13 (MCA2183GA, AbD Serotec, 1:100), Collagen IV (2150-1470, AbD Serotec, 1:100), Desmin (ab15200, Abcam, 1:100), Tbx18 (sc-17869, Santa Cruz, 1:100), αSMA (eBioscience, 50-9760-82, 1:100), oestrogen receptor (ab27595, Abcam, not diluted), β-galactosidase (ab9361, Abcam, 1:100), RFP (600-401-379, Rockland, 1:1,000), and RFP (ABIN334653, ChromoTEK, 1:200).

    Techniques: Mouse Assay, Immunostaining, Clone Assay

    Developmental distribution and molecular properties of PDGFRβ+ cells. ( a – i ) Distribution of PDGFRβ (green) immunostained cells from E10.5 to E14.5. Heart sections from wild-type mice were stained for PDGFRβ (green) and isolectinB4 (blue). Arrows indicate PDGFRβ+ cells, arrowheads mark PDGFRβ- epicardial cells at the indicated stages. Panels at the bottom ( b , c , e , f , h , i ) are higher magnifications of insets in ( a , d , g ), respectively. A, atrium; V, ventricle; AVC, atrioventricular canal; Epi, epicardium; Myo, myocardium; VS, ventricular septum. ( j – l ) Transient co-expression of PDGFRβ (red) and PDGFRα ( Pdgfra tm11(EGFP)Sor reporter; nuclear H2B-GFP; green) during heart development. ECs, isolectinB4 (blue). Arrows indicate GFP+ PDGFRβ+ double-positive cells in the E10.5 AVC ( j ) and E14.5 myocardium ( k ). Double-positive cells were very rare in the E16.5 myocardium ( l ), whereas GFP- PDGFRβ+ mural cells (arrowheads) and GFP+ PDGFRβ- interstitial cells (arrows in l ) were abundant.

    Journal: Nature Communications

    Article Title: Endothelial cells are progenitors of cardiac pericytes and vascular smooth muscle cells

    doi: 10.1038/ncomms12422

    Figure Lengend Snippet: Developmental distribution and molecular properties of PDGFRβ+ cells. ( a – i ) Distribution of PDGFRβ (green) immunostained cells from E10.5 to E14.5. Heart sections from wild-type mice were stained for PDGFRβ (green) and isolectinB4 (blue). Arrows indicate PDGFRβ+ cells, arrowheads mark PDGFRβ- epicardial cells at the indicated stages. Panels at the bottom ( b , c , e , f , h , i ) are higher magnifications of insets in ( a , d , g ), respectively. A, atrium; V, ventricle; AVC, atrioventricular canal; Epi, epicardium; Myo, myocardium; VS, ventricular septum. ( j – l ) Transient co-expression of PDGFRβ (red) and PDGFRα ( Pdgfra tm11(EGFP)Sor reporter; nuclear H2B-GFP; green) during heart development. ECs, isolectinB4 (blue). Arrows indicate GFP+ PDGFRβ+ double-positive cells in the E10.5 AVC ( j ) and E14.5 myocardium ( k ). Double-positive cells were very rare in the E16.5 myocardium ( l ), whereas GFP- PDGFRβ+ mural cells (arrowheads) and GFP+ PDGFRβ- interstitial cells (arrows in l ) were abundant.

    Article Snippet: The following primary antibodies were used: PECAM1 (553370, Pharmingen, 1:100), PDGFRβ (14-1402-82, eBioscience, 1:100), PDGFRα (14-1401-81, eBioscience, 1:100), isolectin B4 (B-1205, Vector, 1:100), NG2 (AB5320, Millipore, 1:100), PDGFRα (3164, Cell Signaling, 1:100), GFP (A21311, Invitrogen, 1:100), GFP (GFP-1010, Aves, 1:200), VE-cadherin (AF1002, R & D Systems, 1:100), CD13 (MCA2183GA, AbD Serotec, 1:100), Collagen IV (2150-1470, AbD Serotec, 1:100), Desmin (ab15200, Abcam, 1:100), Tbx18 (sc-17869, Santa Cruz, 1:100), αSMA (eBioscience, 50-9760-82, 1:100), oestrogen receptor (ab27595, Abcam, not diluted), β-galactosidase (ab9361, Abcam, 1:100), RFP (600-401-379, Rockland, 1:1,000), and RFP (ABIN334653, ChromoTEK, 1:200).

    Techniques: Mouse Assay, Staining, Expressing

    The amino acid dependence of mTOR complex 1 binding to recombinant Rag heterodimers in HeLa cells stably expressing GFP-streptag-RagC variants at differing abundances. HeLa cells stably expressing GFP-streptag, GFP-streptag-RagC WT , or GFP-streptag-RagC S75L were selected by cell sorting for GFP abundance, and the expression of the full-length recombinant protein was estimated by GFP immunoblot analysis of lysates. The cells were incubated in fresh DMEM for 1 h and then deprived of amino acids by incubation in DPBS for 1.5 h. The medium of plates AA- was replaced with fresh DPBS, whereas that of plates AA+ was replaced with DMEM, with harvest 10 min thereafter. Strep-Tactin pull-downs ( PD ) and the lysates were analyzed by immunoblotting for endogenous ( Endo ) Raptor and RagA, for GFP, and for 4E-BP(T37P/T46P).

    Journal: The Journal of Biological Chemistry

    Article Title: Amino Acids Activate Mammalian Target of Rapamycin (mTOR) Complex 1 without Changing Rag GTPase Guanyl Nucleotide Charging *

    doi: 10.1074/jbc.M113.528505

    Figure Lengend Snippet: The amino acid dependence of mTOR complex 1 binding to recombinant Rag heterodimers in HeLa cells stably expressing GFP-streptag-RagC variants at differing abundances. HeLa cells stably expressing GFP-streptag, GFP-streptag-RagC WT , or GFP-streptag-RagC S75L were selected by cell sorting for GFP abundance, and the expression of the full-length recombinant protein was estimated by GFP immunoblot analysis of lysates. The cells were incubated in fresh DMEM for 1 h and then deprived of amino acids by incubation in DPBS for 1.5 h. The medium of plates AA- was replaced with fresh DPBS, whereas that of plates AA+ was replaced with DMEM, with harvest 10 min thereafter. Strep-Tactin pull-downs ( PD ) and the lysates were analyzed by immunoblotting for endogenous ( Endo ) Raptor and RagA, for GFP, and for 4E-BP(T37P/T46P).

    Article Snippet: Anti-β-actin (catalog no. ab8227) and anti-LAMP2 (catalog no. ab25631) were from Abcam. mTOR (7C10) rabbit mAb (catalog no. 2983), raptor (24C12) rabbit mAb (catalog no. 2280), RagA/B (D8B5) rabbit mAb (catalog no. 4357), RagC antibody (catalog no. 3360), S6K [T389P], (1A5) mouse mAb (catalog no. 9206), 4EBP1 phospho-4E-BP1 (Thr-37/46) antibody (catalog no. 9459), GFP antibody (catalog no. 2555), and Myc tag (9B11) mouse mAb (catalog no. 2276) were from Cell Signaling Technology.

    Techniques: Binding Assay, Recombinant, Stable Transfection, Expressing, FACS, Incubation

    The effect of amino acid withdrawal on [ 32 P]guanyl nucleotide content of recombinant Rag heterodimeric complexes associated with lysosomal membranes in 32 P i -labeled HEK293T cells stably expressing recombinant RagA or RagC variants. A , subcellular distribution of endogenous RagC in HEK293E cells. A commercial kit was employed to extract selected subcellular fractions from HEK293E cells. Markers included HSP90 (cytosol, C ), calnexin (endoplasmic reticulum and membranes, M), poly-ADP ribose polymerase ( PARP ) (nucleus, N ), and vimentin (intermediate filaments and cytoskeleton, CS ). The image shows the immunocytochemical localization of endogenous RagC in HeLa cells. Scale bar = 10 μm. RagC, green ; DAPI, blue. B , sucrose density gradient fractionation of HEK293T cells. HEK293T cells were sheared open and fractionated upon a 10-ml 13–60% sucrose density gradient at 100,000 × g for 4.5 h. 500-μl fractions were collected, and 50 μl of each even-numbered fraction ( Fxn ), plus the top fraction ( 1 ), was subjected to SDS-PAGE and analyzed by immunoblotting as indicated. Two exposures are shown for RagA. C , isolation of lysosomes from HeLa cells by differential centrifugation. T , cell homogenate; S1 , supernatant from centrifugation of T at 1000 × g for10 min; P1 , pellet from centrifugation of T at 1000 × g for10 min; S2 , supernatant from centrifugation of S1 at 16,100 × g for 30 min; P2 , the pellet from the latter centrifugation; S3 , the supernatant after centrifugation of S2 at 10 5 × g for 30 min. Each fraction was brought to the volume of the original homogenate, and an equal volume was subjected to SDS-PAGE and analyzed by immunoblotting as indicated. D , the effect of amino acid withdrawal on the [ 32 P]guanyl nucleotide content of stably expressed RagC WT and RagC S75L heterodimeric complexes associated with a LAMP2-containing fraction of 32 Pi-labeled HEK293T cells. Replicate plates containing the GFP-streptag-RagC variants indicated were incubated with 32 P in P i -free DMEM for 4 h, rinsed, and incubated in either homemade P i -free medium ( AA +) or P i -free medium lacking amino acids ( AA -), each containing 32 P i as in Fig. 6 . The cells were extracted two h later, the fraction corresponding to P2 ( C ) was isolated, and the [ 32 P]guanyl nucleotide content of Strep-Tactin-isolated Rag complexes in P2 was determined as in Fig. 6 . Immunoblot analyses of GFP in the P2 fraction and 4E-BP(T37P/T46P) in the lysate are shown in the right panel. E , the effect of amino acid withdrawal on the [ 32 P]guanyl nucleotide content of stably expressed RagA WT and RagA T21L heterodimeric complexes associated with a LAMP2-containing fraction of 32 Pi-labeled HEK293T cells. The cells and extracts were processed as described in D . Immunoblots of GFP and RagA in the P2 fraction and 4E-BPT37P/T46P) in the lysates are shown in the right panel. Ori , origin; Ct , C-terminal.

    Journal: The Journal of Biological Chemistry

    Article Title: Amino Acids Activate Mammalian Target of Rapamycin (mTOR) Complex 1 without Changing Rag GTPase Guanyl Nucleotide Charging *

    doi: 10.1074/jbc.M113.528505

    Figure Lengend Snippet: The effect of amino acid withdrawal on [ 32 P]guanyl nucleotide content of recombinant Rag heterodimeric complexes associated with lysosomal membranes in 32 P i -labeled HEK293T cells stably expressing recombinant RagA or RagC variants. A , subcellular distribution of endogenous RagC in HEK293E cells. A commercial kit was employed to extract selected subcellular fractions from HEK293E cells. Markers included HSP90 (cytosol, C ), calnexin (endoplasmic reticulum and membranes, M), poly-ADP ribose polymerase ( PARP ) (nucleus, N ), and vimentin (intermediate filaments and cytoskeleton, CS ). The image shows the immunocytochemical localization of endogenous RagC in HeLa cells. Scale bar = 10 μm. RagC, green ; DAPI, blue. B , sucrose density gradient fractionation of HEK293T cells. HEK293T cells were sheared open and fractionated upon a 10-ml 13–60% sucrose density gradient at 100,000 × g for 4.5 h. 500-μl fractions were collected, and 50 μl of each even-numbered fraction ( Fxn ), plus the top fraction ( 1 ), was subjected to SDS-PAGE and analyzed by immunoblotting as indicated. Two exposures are shown for RagA. C , isolation of lysosomes from HeLa cells by differential centrifugation. T , cell homogenate; S1 , supernatant from centrifugation of T at 1000 × g for10 min; P1 , pellet from centrifugation of T at 1000 × g for10 min; S2 , supernatant from centrifugation of S1 at 16,100 × g for 30 min; P2 , the pellet from the latter centrifugation; S3 , the supernatant after centrifugation of S2 at 10 5 × g for 30 min. Each fraction was brought to the volume of the original homogenate, and an equal volume was subjected to SDS-PAGE and analyzed by immunoblotting as indicated. D , the effect of amino acid withdrawal on the [ 32 P]guanyl nucleotide content of stably expressed RagC WT and RagC S75L heterodimeric complexes associated with a LAMP2-containing fraction of 32 Pi-labeled HEK293T cells. Replicate plates containing the GFP-streptag-RagC variants indicated were incubated with 32 P in P i -free DMEM for 4 h, rinsed, and incubated in either homemade P i -free medium ( AA +) or P i -free medium lacking amino acids ( AA -), each containing 32 P i as in Fig. 6 . The cells were extracted two h later, the fraction corresponding to P2 ( C ) was isolated, and the [ 32 P]guanyl nucleotide content of Strep-Tactin-isolated Rag complexes in P2 was determined as in Fig. 6 . Immunoblot analyses of GFP in the P2 fraction and 4E-BP(T37P/T46P) in the lysate are shown in the right panel. E , the effect of amino acid withdrawal on the [ 32 P]guanyl nucleotide content of stably expressed RagA WT and RagA T21L heterodimeric complexes associated with a LAMP2-containing fraction of 32 Pi-labeled HEK293T cells. The cells and extracts were processed as described in D . Immunoblots of GFP and RagA in the P2 fraction and 4E-BPT37P/T46P) in the lysates are shown in the right panel. Ori , origin; Ct , C-terminal.

    Article Snippet: Anti-β-actin (catalog no. ab8227) and anti-LAMP2 (catalog no. ab25631) were from Abcam. mTOR (7C10) rabbit mAb (catalog no. 2983), raptor (24C12) rabbit mAb (catalog no. 2280), RagA/B (D8B5) rabbit mAb (catalog no. 4357), RagC antibody (catalog no. 3360), S6K [T389P], (1A5) mouse mAb (catalog no. 9206), 4EBP1 phospho-4E-BP1 (Thr-37/46) antibody (catalog no. 9459), GFP antibody (catalog no. 2555), and Myc tag (9B11) mouse mAb (catalog no. 2276) were from Cell Signaling Technology.

    Techniques: Recombinant, Labeling, Stable Transfection, Expressing, Fractionation, SDS Page, Isolation, Centrifugation, Incubation, Western Blot

    The effect of amino acid withdrawal and insulin on the [ 32 P]guanyl nucleotide content of stably expressed RagC WT and RagC S75L heterodimeric complexes in 32 P i -labeled HeLa cells. A , B , and C , replicate plates of HeLa cells stably expressing GFP-streptag, GFP-streptag-RagC WT , or GFP-streptag-RagC S75L were incubated in P i -free DMEM containing 32 P i (0.2 mCi/ml). After 4 h, the cells were rinsed and incubated in either homemade P i -free medium ( AA +) or P i -free medium lacking amino acids ( AA -), each containing 32 P i (0.2 mCi/ml), for another 2 h. In A and B , insulin (1.0 μ m ) was added to some of the cells in DMEM ( AA +/ I ) 30 min before harvest. The nucleotides bound to Strep-Tactin ( strept ) pull-downs were extracted and separated by TLC on PEI cellulose. One set of 32 P -labeled HeLa cells expressing FP-streptag, treated as described above for AA-, AA+, and AA+/I, were rinsed, extracted directly into acetonitrile, and then the solubilized total nucleotides were separated by TLC. The 32 P comigrating with GDP and GTP was quantitated by phosphorimaging. After subtraction of the averaged values found in the GFP-streptag lanes, the percentage of [ 32 P]GTP was calculated as [[ 32 P]GTP / (1.5 × [ 32 P]GDP + [ 32 P]GTP)], and the averaged values are shown. In A , the top and center panels on the right show GFP immunoblot analyses of the cell lysates and representative Strep-Tactin pull-downs (corresponding to lanes 1 , 4 , and 7 of each set). The bottom panel shows an immunoblot analysis of S6K(T389P) corresponding to lanes 1 , 4 , and 7 of each set. In B , the top panel on the right shows a GFP immunoblot analysis of the lysates, whereas the center and bottom panels show lysate immunoblot analyses for 4E-BP(T37P/T46P) and PRAS40(S246P), respectively. In C , one set of 32 P-labeled HeLa cells expressing GFP-streptag, treated as in Fig. 6 , were rinsed, extracted directly into HClO 4 (0.3 m , 0 °C, HClO 4 extract ). The HClO 4 supernatants were neutralized with KHCO 3 , and the [ 32 P]guanyl nucleotides were quantified as above. In addition, the nucleotides in the lysate after pull-down of the Strep-Tactin beads were also analyzed. Immunoblot analyses of the lysates and representative Strep-Tactin pull-downs are shown in the right panel. Ori , origin; std , guanyl nucleotide standards; Ins , insulin; ACN , acetonitrile; Ct , C-terminal.

    Journal: The Journal of Biological Chemistry

    Article Title: Amino Acids Activate Mammalian Target of Rapamycin (mTOR) Complex 1 without Changing Rag GTPase Guanyl Nucleotide Charging *

    doi: 10.1074/jbc.M113.528505

    Figure Lengend Snippet: The effect of amino acid withdrawal and insulin on the [ 32 P]guanyl nucleotide content of stably expressed RagC WT and RagC S75L heterodimeric complexes in 32 P i -labeled HeLa cells. A , B , and C , replicate plates of HeLa cells stably expressing GFP-streptag, GFP-streptag-RagC WT , or GFP-streptag-RagC S75L were incubated in P i -free DMEM containing 32 P i (0.2 mCi/ml). After 4 h, the cells were rinsed and incubated in either homemade P i -free medium ( AA +) or P i -free medium lacking amino acids ( AA -), each containing 32 P i (0.2 mCi/ml), for another 2 h. In A and B , insulin (1.0 μ m ) was added to some of the cells in DMEM ( AA +/ I ) 30 min before harvest. The nucleotides bound to Strep-Tactin ( strept ) pull-downs were extracted and separated by TLC on PEI cellulose. One set of 32 P -labeled HeLa cells expressing FP-streptag, treated as described above for AA-, AA+, and AA+/I, were rinsed, extracted directly into acetonitrile, and then the solubilized total nucleotides were separated by TLC. The 32 P comigrating with GDP and GTP was quantitated by phosphorimaging. After subtraction of the averaged values found in the GFP-streptag lanes, the percentage of [ 32 P]GTP was calculated as [[ 32 P]GTP / (1.5 × [ 32 P]GDP + [ 32 P]GTP)], and the averaged values are shown. In A , the top and center panels on the right show GFP immunoblot analyses of the cell lysates and representative Strep-Tactin pull-downs (corresponding to lanes 1 , 4 , and 7 of each set). The bottom panel shows an immunoblot analysis of S6K(T389P) corresponding to lanes 1 , 4 , and 7 of each set. In B , the top panel on the right shows a GFP immunoblot analysis of the lysates, whereas the center and bottom panels show lysate immunoblot analyses for 4E-BP(T37P/T46P) and PRAS40(S246P), respectively. In C , one set of 32 P-labeled HeLa cells expressing GFP-streptag, treated as in Fig. 6 , were rinsed, extracted directly into HClO 4 (0.3 m , 0 °C, HClO 4 extract ). The HClO 4 supernatants were neutralized with KHCO 3 , and the [ 32 P]guanyl nucleotides were quantified as above. In addition, the nucleotides in the lysate after pull-down of the Strep-Tactin beads were also analyzed. Immunoblot analyses of the lysates and representative Strep-Tactin pull-downs are shown in the right panel. Ori , origin; std , guanyl nucleotide standards; Ins , insulin; ACN , acetonitrile; Ct , C-terminal.

    Article Snippet: Anti-β-actin (catalog no. ab8227) and anti-LAMP2 (catalog no. ab25631) were from Abcam. mTOR (7C10) rabbit mAb (catalog no. 2983), raptor (24C12) rabbit mAb (catalog no. 2280), RagA/B (D8B5) rabbit mAb (catalog no. 4357), RagC antibody (catalog no. 3360), S6K [T389P], (1A5) mouse mAb (catalog no. 9206), 4EBP1 phospho-4E-BP1 (Thr-37/46) antibody (catalog no. 9459), GFP antibody (catalog no. 2555), and Myc tag (9B11) mouse mAb (catalog no. 2276) were from Cell Signaling Technology.

    Techniques: Stable Transfection, Labeling, Expressing, Incubation, Thin Layer Chromatography

    Ectopic expression of HNF6 in acinar cells is associated with induction of ductal genes and repression of acinar genes in vitro . (A) Relative expression of ductal (left) and acinar (right) genes in cultured 266-6 cells. Cells were transduced with an adenovirus expressing GFP (Ad-GFP), HNF6 (Ad-HNF6), or Sox9 (Ad-Sox9) and gene expression was analyzed by Q-RT-PCR (data are means +/− SEM; n= 4; *, p

    Journal: Gut

    Article Title: Role of the ductal transcription factors HNF6 and Sox9 in pancreatic acinar-to-ductal metaplasia

    doi: 10.1136/gutjnl-2011-300266

    Figure Lengend Snippet: Ectopic expression of HNF6 in acinar cells is associated with induction of ductal genes and repression of acinar genes in vitro . (A) Relative expression of ductal (left) and acinar (right) genes in cultured 266-6 cells. Cells were transduced with an adenovirus expressing GFP (Ad-GFP), HNF6 (Ad-HNF6), or Sox9 (Ad-Sox9) and gene expression was analyzed by Q-RT-PCR (data are means +/− SEM; n= 4; *, p

    Article Snippet: Cells were infected with adenovirus expressing either HNF6 (Ad-HNF6), Sox9 (Ad-Sox9),[ ] or GFP (Ad-GFP) (Vectorbiolabs) at a multiplicity of infection of 100 in 200 μl of DMEM.

    Techniques: Expressing, In Vitro, Cell Culture, Transduction, Reverse Transcription Polymerase Chain Reaction

    Ectopic expression of HNF6 in acinar cells is associated with induction of ductal genes and repression of acinar genes in vivo . (A–D) Immunolabeling and immunohistochemical staining of sections of wild-type (A–C) and ElaC-Sox9 f/f (D) pancreata 3 days after infection by Ad-GFP (A), Ad-Sox9 (B), or Ad-HNF6 (C,D). Metaplasia is detected in wild-type pancreas infected with Ad-HNF6 (C). Ad-HNF6 induces Sox9, and induces the ductal marker CK19 in the cytoplasm of acinar cells, while repressing Amylase. In acinar cells, Ad-HNF6 relocates Ezrin from the apical pole to the cytoplasm. After Ad-Sox9 infection, HNF6, Amylase and Ezrin are unaffected, but marginal induction of CK19 is detectable in acinar cells (B). No induction of Sox9 is observed in ElaC-Sox9 f/f after infection with Ad-HNF6 (D). Ad-HNF6 repressed Amylase in ElaC-Sox9 f/f ). (E) Percentage of GFP-positive cells that coexpressed HNF6 or Sox9, two (green) or three (yellow) days after Ad-HNF6 infection (left panel). Quantification of the percentage of Sox9-positive cells that coexpressed HNF6 at the same time points. These quantifications show that HNF6 expression is less stable than GFP and Sox9 (right panel). (F) GFP/HNF6 immunolabeling of pancreas two or three days after transduction with Ad-HNF6, which codes for both HNF6 and GFP. HNF6 is less stably expressed than GFP. (G) Cleaved Caspase3/GFP and Cleaved Caspase3/HNF6 immunolabelling of pancreas infected by Ad-GFP and Ad-HNF6, respectively. Many apoptotic cells are detected in pancreas infected with Ad-HNF6. Cl. Caspase3, Cleaved Caspase3.

    Journal: Gut

    Article Title: Role of the ductal transcription factors HNF6 and Sox9 in pancreatic acinar-to-ductal metaplasia

    doi: 10.1136/gutjnl-2011-300266

    Figure Lengend Snippet: Ectopic expression of HNF6 in acinar cells is associated with induction of ductal genes and repression of acinar genes in vivo . (A–D) Immunolabeling and immunohistochemical staining of sections of wild-type (A–C) and ElaC-Sox9 f/f (D) pancreata 3 days after infection by Ad-GFP (A), Ad-Sox9 (B), or Ad-HNF6 (C,D). Metaplasia is detected in wild-type pancreas infected with Ad-HNF6 (C). Ad-HNF6 induces Sox9, and induces the ductal marker CK19 in the cytoplasm of acinar cells, while repressing Amylase. In acinar cells, Ad-HNF6 relocates Ezrin from the apical pole to the cytoplasm. After Ad-Sox9 infection, HNF6, Amylase and Ezrin are unaffected, but marginal induction of CK19 is detectable in acinar cells (B). No induction of Sox9 is observed in ElaC-Sox9 f/f after infection with Ad-HNF6 (D). Ad-HNF6 repressed Amylase in ElaC-Sox9 f/f ). (E) Percentage of GFP-positive cells that coexpressed HNF6 or Sox9, two (green) or three (yellow) days after Ad-HNF6 infection (left panel). Quantification of the percentage of Sox9-positive cells that coexpressed HNF6 at the same time points. These quantifications show that HNF6 expression is less stable than GFP and Sox9 (right panel). (F) GFP/HNF6 immunolabeling of pancreas two or three days after transduction with Ad-HNF6, which codes for both HNF6 and GFP. HNF6 is less stably expressed than GFP. (G) Cleaved Caspase3/GFP and Cleaved Caspase3/HNF6 immunolabelling of pancreas infected by Ad-GFP and Ad-HNF6, respectively. Many apoptotic cells are detected in pancreas infected with Ad-HNF6. Cl. Caspase3, Cleaved Caspase3.

    Article Snippet: Cells were infected with adenovirus expressing either HNF6 (Ad-HNF6), Sox9 (Ad-Sox9),[ ] or GFP (Ad-GFP) (Vectorbiolabs) at a multiplicity of infection of 100 in 200 μl of DMEM.

    Techniques: Expressing, In Vivo, Immunolabeling, Immunohistochemistry, Staining, Infection, Marker, Transduction, Stable Transfection

    GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using a single-photon counting method. ( A ) Steady-state intensity image of GFP-BD expressed alone in HEK293 cell ( green ). The corresponding GFP-BD fluorescence decay ( green

    Journal:

    Article Title: Quantitative FRET Analysis by Fast Acquisition Time Domain FLIM at High Spatial Resolution in Living Cells

    doi: 10.1529/biophysj.108.131276

    Figure Lengend Snippet: GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using a single-photon counting method. ( A ) Steady-state intensity image of GFP-BD expressed alone in HEK293 cell ( green ). The corresponding GFP-BD fluorescence decay ( green

    Article Snippet: We tagged the TAFII 250 double BD module with GFP (GFP-BD) and the histone H4 with the red fluorescent protein, mCherry-H4 (mCherry) to quantify and visualize histone acetylation in living cells by FRET imaging.

    Techniques: Fluorescence

    FLIM and mf D images of GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using TriM-FLIM system at fast acquisition times. ( A ) Intensity and FLIM images of GFP-BD expressed alone as control ( upper panel ) or with mCherry-H4

    Journal:

    Article Title: Quantitative FRET Analysis by Fast Acquisition Time Domain FLIM at High Spatial Resolution in Living Cells

    doi: 10.1529/biophysj.108.131276

    Figure Lengend Snippet: FLIM and mf D images of GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using TriM-FLIM system at fast acquisition times. ( A ) Intensity and FLIM images of GFP-BD expressed alone as control ( upper panel ) or with mCherry-H4

    Article Snippet: We tagged the TAFII 250 double BD module with GFP (GFP-BD) and the histone H4 with the red fluorescent protein, mCherry-H4 (mCherry) to quantify and visualize histone acetylation in living cells by FRET imaging.

    Techniques:

    FLIM, f D and mf D images of GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using the TriM-FLIM system with 11 time-gated images. ( A ) Intensity, FLIM, f D , and mf D images of GFP-BD expressed alone as control ( upper panel

    Journal:

    Article Title: Quantitative FRET Analysis by Fast Acquisition Time Domain FLIM at High Spatial Resolution in Living Cells

    doi: 10.1529/biophysj.108.131276

    Figure Lengend Snippet: FLIM, f D and mf D images of GFP-BD interaction with acetylated mCherry-H4 in the nucleus of HEK293 live cells using the TriM-FLIM system with 11 time-gated images. ( A ) Intensity, FLIM, f D , and mf D images of GFP-BD expressed alone as control ( upper panel

    Article Snippet: We tagged the TAFII 250 double BD module with GFP (GFP-BD) and the histone H4 with the red fluorescent protein, mCherry-H4 (mCherry) to quantify and visualize histone acetylation in living cells by FRET imaging.

    Techniques:

    Mitochondria are transferred from iPSC-MSCs to injured PC12 cells under CoCl 2 challenge. (A) TNTs formed between PC12 cells (in green) and iPSC-MSCs. Scale bar: 10 μm. (B) The number of iPSC-MSC TNTs was greatly enhanced under CoCl 2 challenge. (C) Mitochondria were transferred from Mito-GFP-iPSC-MSCs (in green) to Celltrace-labelled-PC12 cells (in blue) via TNTs. Scale bar: 10 μm. (D) The efficiency of mitochondrial transfer from iPSC-MSCs to PC12 cells was dramatically enhanced under CoCl 2 treatment compared with no CoCl 2 treatment. (E) ATP levels were dramatically increased in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. (F) Apoptosis was dramatically reduced in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. Data are expressed as the mean ± SEM ( n = 3; unpaired Student’s t -test). All experiments were conducted in triplicate. * P

    Journal: Neural Regeneration Research

    Article Title: Transfer of mitochondria from mesenchymal stem cells derived from induced pluripotent stem cells attenuates hypoxia-ischemia-induced mitochondrial dysfunction in PC12 cells

    doi: 10.4103/1673-5374.266058

    Figure Lengend Snippet: Mitochondria are transferred from iPSC-MSCs to injured PC12 cells under CoCl 2 challenge. (A) TNTs formed between PC12 cells (in green) and iPSC-MSCs. Scale bar: 10 μm. (B) The number of iPSC-MSC TNTs was greatly enhanced under CoCl 2 challenge. (C) Mitochondria were transferred from Mito-GFP-iPSC-MSCs (in green) to Celltrace-labelled-PC12 cells (in blue) via TNTs. Scale bar: 10 μm. (D) The efficiency of mitochondrial transfer from iPSC-MSCs to PC12 cells was dramatically enhanced under CoCl 2 treatment compared with no CoCl 2 treatment. (E) ATP levels were dramatically increased in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. (F) Apoptosis was dramatically reduced in the PC12 cells that received mitochondria from iPSC-MSCs compared with those that did not. Data are expressed as the mean ± SEM ( n = 3; unpaired Student’s t -test). All experiments were conducted in triplicate. * P

    Article Snippet: Mitochondrial transfer assessment To examine mitochondrial transfer, iPSC-MSCs were infected with lentivirus-containing mitochondria with GFP (mito-GFP) (Cyto102-PA-1, System Biosciences, Palo Alto, CA, USA), as previously described (Zhang et al., 2016).

    Techniques:

    Immunofluorescent stainings against GABA . GABA immunoreactivity shown alone in gray (A2,B2,C2,D2) , or (in magenta) overlay with Ccap-Gal4-driven expression of GFP (green, A1,B1,C1,D1 ). In the brain [ (A1,A2) , projection of 8 confocal slices containing the IN brain ] and the sog [ (B1,B2) , projection of 2 confocal slices containing the IN sog ], all N CCAP are GABA-immunonegative. In some but not all preparations of the thoracic and abdominal neuromeres, GABA immunoreactivity was found in one N CCAP per hemineuromere. (C1,C2) While the N CCAP on the right side of a4 and a5 are GABA-immunonegative (arrowheads), there is one N CCAP on the left side in a5 that shows weak but distinct GABA labeling (arrow, projection of 2 confocal slices). (D1,D2) shows a projection of 8 confocal slices of another preparation, where the N CCAP in a1, a3 and the anterior N CCAP in a4 appear to be GABA-immunonegative (upper arrows). One of the N CCAP in a4 (lowermost arrow) however shows weak GABA immunoreactivity. Scale bars = 50 μm.

    Journal: Frontiers in Neural Circuits

    Article Title: Diverse in- and output polarities and high complexity of local synaptic and non-synaptic signaling within a chemically defined class of peptidergic Drosophila neurons

    doi: 10.3389/fncir.2013.00127

    Figure Lengend Snippet: Immunofluorescent stainings against GABA . GABA immunoreactivity shown alone in gray (A2,B2,C2,D2) , or (in magenta) overlay with Ccap-Gal4-driven expression of GFP (green, A1,B1,C1,D1 ). In the brain [ (A1,A2) , projection of 8 confocal slices containing the IN brain ] and the sog [ (B1,B2) , projection of 2 confocal slices containing the IN sog ], all N CCAP are GABA-immunonegative. In some but not all preparations of the thoracic and abdominal neuromeres, GABA immunoreactivity was found in one N CCAP per hemineuromere. (C1,C2) While the N CCAP on the right side of a4 and a5 are GABA-immunonegative (arrowheads), there is one N CCAP on the left side in a5 that shows weak but distinct GABA labeling (arrow, projection of 2 confocal slices). (D1,D2) shows a projection of 8 confocal slices of another preparation, where the N CCAP in a1, a3 and the anterior N CCAP in a4 appear to be GABA-immunonegative (upper arrows). One of the N CCAP in a4 (lowermost arrow) however shows weak GABA immunoreactivity. Scale bars = 50 μm.

    Article Snippet: Immunostainings and GFP-labeling CNS from third instar larvae were dissected in standard fly saline or PBS, fixed for 45 min to 4 h in 4% paraformaldehyde in 0.1 M sodium phosphate buffered saline (PBS, pH 7.2) at 4°C, washed in PBS with 1% TritonX (PBT) and incubated for at least 24 h in PBT containing 10% normal goat serum in combination with rabbit polyclonal (anti-PRXa (Eckert et al., , 1:5000), anti-CCAP (Dircksen and Keller, , 1:1000), anti-GABA (Sigma-Aldrich, 1:800, Thum et al., ), a-GFP (Invitrogen, 1:1000) or mouse monoclonal antibodies (a-brp nc82 (1:100), anti-GFP (Invitrogen) 1:1000), as well as a-FasII 1D4 (1:75), and a-ChAT 4B1 (1:50) obtained from the Developmental Studies Hybridoma Bank under the auspices of the NICHD and maintained by the University of Iowa.

    Techniques: Expressing, Labeling

    BAY 11-7082 suppresses the LPS- or IL-1-stimulated formation of K63-pUb chains and the DNA damage response ( A and B ) The experiment was carried out as in Figure 2 , except that the K63-pUb chains formed in response to LPS ( A ) or IL-1β ( B ) were captured on Halo-NEMO from 6 mg (RAW cells) or 3 mg (IL-1R cells) of cell extract protein as described in the Experimental section. The K63-pUb chains were identified by immunoblotting with a specific antibody. Further aliquots of the cell extract were immunoblotted for IKKβ phosphorylation, p105 phosphorylation and GAPDH as in Figure 2 . ( C ) Indirect immunofluorescence images of U2OS cells transiently expressing GFP–RAP80[1–200]. Cells were incubated for 1 h with or without BAY 11-7082 (15 μM) and either exposed to ionizing radiation (IR) or not exposed. GFP–RAP80 or γH2AX were visualized using anti-GFP and anti-γH2AX antibodies respectively, and nuclei were stained with DAPI.

    Journal: Biochemical Journal

    Article Title: The anti-inflammatory drug BAY 11-7082 suppresses the MyD88-dependent signalling network by targeting the ubiquitin system

    doi: 10.1042/BJ20121651

    Figure Lengend Snippet: BAY 11-7082 suppresses the LPS- or IL-1-stimulated formation of K63-pUb chains and the DNA damage response ( A and B ) The experiment was carried out as in Figure 2 , except that the K63-pUb chains formed in response to LPS ( A ) or IL-1β ( B ) were captured on Halo-NEMO from 6 mg (RAW cells) or 3 mg (IL-1R cells) of cell extract protein as described in the Experimental section. The K63-pUb chains were identified by immunoblotting with a specific antibody. Further aliquots of the cell extract were immunoblotted for IKKβ phosphorylation, p105 phosphorylation and GAPDH as in Figure 2 . ( C ) Indirect immunofluorescence images of U2OS cells transiently expressing GFP–RAP80[1–200]. Cells were incubated for 1 h with or without BAY 11-7082 (15 μM) and either exposed to ionizing radiation (IR) or not exposed. GFP–RAP80 or γH2AX were visualized using anti-GFP and anti-γH2AX antibodies respectively, and nuclei were stained with DAPI.

    Article Snippet: Antibodies that recognize GFP (green fluorescent protein) (Abcam), K63-pUb chains (eBioscience), K48-pUb chains, IRAK4 and histone γH2AX (Merck-Millipore) were purchased from the sources indicated.

    Techniques: Immunofluorescence, Expressing, Incubation, Staining

    Effect of AAV2.5-driven SERCA2a gene transfer on post-injury vascula healing Two groups of animals were analyzed: sham-operated control, injured and AAV2.5-GFP (n=10) or AAV2.5-SERCA2a (n=8) infected carotid artery 1 month after surgery. A . Representative hematoxylin/eosin staining of carotid artery cross-section. Objective X10 (upper panel), X60 (lower panel); ni – neointima; m – media; a – adventitia, lm -lumen. B . Morphometric analysis of carotid artery cross-sections. Bars represent the mean ± SEM of mean values obtained for each animal. At least 5 individual measures were performed for each animal on different carotid cross sections.

    Journal: Gene therapy

    Article Title: Efficient transduction of vascular smooth muscle cells with a translational AAV2.5 vector: a new perspective for in-stent restenosis gene therapy

    doi: 10.1038/gt.2013.13

    Figure Lengend Snippet: Effect of AAV2.5-driven SERCA2a gene transfer on post-injury vascula healing Two groups of animals were analyzed: sham-operated control, injured and AAV2.5-GFP (n=10) or AAV2.5-SERCA2a (n=8) infected carotid artery 1 month after surgery. A . Representative hematoxylin/eosin staining of carotid artery cross-section. Objective X10 (upper panel), X60 (lower panel); ni – neointima; m – media; a – adventitia, lm -lumen. B . Morphometric analysis of carotid artery cross-sections. Bars represent the mean ± SEM of mean values obtained for each animal. At least 5 individual measures were performed for each animal on different carotid cross sections.

    Article Snippet: Confocal Immunofluorescence Immunostaining was performed using the following primary antibodies: a-GFP (Abcam), a-SERCA2a ; a-cyclin D1 (556470,BD Biosciences); a-CD31 (Abcam); a-SMMS, s mooth m uscle m yosin heavy chain s 1 and 2 (Abcam); a-eNOS, e ndothelial n itric o xide s ynthase (Abcam) and secondary antibodies conjugated to Alexa-546 or Alexa-488.

    Techniques: Infection, Staining