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  • 88
    TaKaRa morange tag
    Confocal images showing localization of <t>GFP:TGBp2</t> with actin, Golgi, and localization of GFP:TGBp1. A to C, Cells expressing GFP:TGBp2 and <t>DsRed:Talin.</t> Arrows point to vesicles along actin filaments. D to F, Cells expressing GFP:TGBp2 and GFP:Talin. Yellow arrowheads point to vesicles. F, Vesicles in cells treated with latrunculin B. G to J, Tobacco epidermal cells cobombarded with GFP-TGBp2 and DsRed-ST. Note that the GFP-TGBp2 vesicles (arrowhead) do not colocalize with the DsRed-ST decorated Golgi (arrows). K, Cells expressing GFP:TGBp2. L, Cells expressing GFP:TGBp1. Bars represent 10 μ m.
    Morange Tag, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Addgene inc hsyn quasar2 morange
    Development and characterization of QuasAr3. (a) Screening pipeline. Rationally designed constructs were cloned in the Optopatch configuration, expressed in primary neurons and tested for spike SNR using light-induced spikes. Constructs with improved SNR were then expressed in vivo using in utero electroporation (IUE) and tested for spike SNR in acute slices. The process was repeated iteratively. (b) Examples of SNR measurements in cultured neurons. Left: wide-field epifluorescence images of GFP fused to CheRiff, an opsin with excellent membrane trafficking. Middle: Fluorescence of QuasAr mutants. Scale bar 10 μm. Right: QuasAr fluorescence transients in response to optogenetically induced spikes (10 ms blue light stimulation at 1 mW/mm 2 ). Each construct was tested on at least 5 cultured neurons. (c) Hierarchical screen for improved membrane trafficking of QuasAr variants (see Methods for details). Diagram: schematic of the FCK_DuEx1.0 construct and overview of the screening pipeline. E. coli colonies were transformed with libraries in FCK_DuEx1.0. The colonies with the brightest fluorescence were picked for lentivirus production and secondary screening in primary neuronal culture. Images: example images of the FP channel of <t>QuasAr2-FP</t> fusions: i. <t>mOrange;</t> ii. mRuby2; iii. mKate2; iv. Citrine. Scale bar 10 μm. (d) SNR of N-terminal modifications compared with QuasAr2. All constructs showed reduced SNR (see Methods for details). (e) Replacing mOrange2 with Citrine as a fusion protein improved the trafficking only with two specific linkers. (f) Adding additional TS sequences at the linker and C terminal improved the spike SNR. (g) The mutation K171R increased the QuasAr expression level, quantified by normalizing QuasAr fluorescence by the fluorescence of the co-expressed CheRiff-GFP. (d-g) all error bars are mean ± s.e.m., 1-tail t -test. (h) Top: diagram of the QuasAr2 and QuasAr3 constructs. Bottom: Confocal images of brain slices expressing QuasAr2 and QuasAr3. Scale bar 500 μm. Insets: single cell bodies, scale bar 10 μm. Representative images from N = 2 mice (QuasAr2) and N = 3 mice (QuasAr3). (i) Confocal images of brain slices expressing Cre-dependent QuasAr3 with sparsity controlled by co-expression of <t>hSyn-Cre.</t> Scale bar 50 μm. (j) Simultaneous fluorescence and patch clamp recordings from two neurons expressing QuasAr3 using AAV virus in acute brain slice. Left: image of QuasAr3 fluorescence in the soma. Scale bar 10 μm. Middle: spiking during ramp current injection. Right: mean spike, overlay of fluorescence and voltage. Inset: boxed regions showing correspondence of optical and electrical recordings of sub-threshold voltage overlaid. See Extended Data Fig. 3 for statistics. Scale bar 10 μm.
    Hsyn Quasar2 Morange, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Chroma Technology Corporation dsred
    <t>DsRed</t> fluorescence imaging (A) shows the location of 4T1.2neu/R primary tumor in Balb/c mice, 2 weeks after subcutaneous inoculation into the mammary pad. <t>NIR</t> fluorescence (B) and PET/CT (C) imaging performed 24 hours after intravenous administration
    Dsred, supplied by Chroma Technology Corporation, used in various techniques. Bioz Stars score: 92/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Omega Optical dsred
    Fig. 4. In vivo localization of <t>Hmo1–GFP</t> and Fob1–GFP fusion proteins. ( A ) were grown at 30°C to mid-log phase (OD 600 = 0.5) in YPD. Cells were washed with water. Nucleoplasm and nucleolus were visualized using DNA staining and Nop1 staining, respectively. GFP (green), <t>DsRed</t> (red) and Hoechst 33352 (blue) signals were monitored by fluorescence microscopy as described in Materials and methods. Cells were examined by Nomarski imaging. ( B ) Fob1 and Hmo1 co-localize. Fob1 and Hmo1 were tagged with different GFP spectral variants, CFP and YFP, and visualized in a diploid strain prepared by crossing strains BMA-Fob1–CFP and BMA-HMO1–YFP as described in Materials and methods (scale bars correspond to 2 µm).
    Dsred, supplied by Omega Optical, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson dsred
    125–267, which contains TMD4R but lacks TMD2R, only localizes to peroxisomes in uninduced cells. Colocalization of <t>Pmp47-GFP</t> fusion proteins and <t>DsRed-AKL</t> is shown. (A) Representative single cells from glucose and glycerol cultures. (B) Representative cells from oleate cultures.
    Dsred, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Carl Zeiss dsred
    Effect of TIGS of candidate genes on the number of <t>DsRed</t> fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and <t>GFP</t> expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P
    Dsred, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 92/100, based on 159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher dsred
    Effect of TIGS of candidate genes on the number of <t>DsRed</t> fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and <t>GFP</t> expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P
    Dsred, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    AHF analysentechnik dsred
    Effect of TIGS of candidate genes on the number of <t>DsRed</t> fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and <t>GFP</t> expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P
    Dsred, supplied by AHF analysentechnik, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa dsred c1 dsred
    Effect of TIGS of candidate genes on the number of <t>DsRed</t> fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and <t>GFP</t> expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P
    Dsred C1 Dsred, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    The Jackson Laboratory dsred
    Wild-type mouse HSPC-derived cells deliver frataxin and Cox8 to FRDA cells in vitro and in vivo ( A and B ). Scale bars, 10 μm. ( C ) Representative confocal images of brain sections from a YG8R mouse transplanted with <t>DsRed</t> + ) and those of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + <t>/Cox8-GFP</t> + for 3D visualization). Scale bars, 10 μm. ( D ) Representative confocal images of a spinal cord section from a YG8R mouse transplanted with DsRed + /Cox8-GFP + HSPCs at 7 months after transplantation, labeled with an anti-NeuN antibody (blue). White arrows depict Cox8-GFP within branch extensions of the DsRed + microglial cell. Scale bar, 5 μm. ( E ) Quantification of neurons containing Cox8-GFP in the cervical spinal cord gray matter of YG8R mice transplanted with DsRed + /Cox8-GFP + for the description of the automatic unbiased quantification method). ( F ) Representative confocal images of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + HSPCs transduced with LV-hFXN-GFP at 7 months after transplantation and stained with anti-mCherry (red) and anti-NeuN (blue) antibodies. Scale bars, 10 μm.
    Dsred, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 92/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Leica Microsystems dsred
    Wild-type mouse HSPC-derived cells deliver frataxin and Cox8 to FRDA cells in vitro and in vivo ( A and B ). Scale bars, 10 μm. ( C ) Representative confocal images of brain sections from a YG8R mouse transplanted with <t>DsRed</t> + ) and those of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + <t>/Cox8-GFP</t> + for 3D visualization). Scale bars, 10 μm. ( D ) Representative confocal images of a spinal cord section from a YG8R mouse transplanted with DsRed + /Cox8-GFP + HSPCs at 7 months after transplantation, labeled with an anti-NeuN antibody (blue). White arrows depict Cox8-GFP within branch extensions of the DsRed + microglial cell. Scale bar, 5 μm. ( E ) Quantification of neurons containing Cox8-GFP in the cervical spinal cord gray matter of YG8R mice transplanted with DsRed + /Cox8-GFP + for the description of the automatic unbiased quantification method). ( F ) Representative confocal images of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + HSPCs transduced with LV-hFXN-GFP at 7 months after transplantation and stained with anti-mCherry (red) and anti-NeuN (blue) antibodies. Scale bars, 10 μm.
    Dsred, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Addgene inc dsred
    Sirt4-sfGFP is exclusively located at the outer mitochondrial membrane. ( a ) Representative SIM images of HeLa cells expressing Sirt4-sfGFP (green, left panel) and mCherry-translocase of the outer mitochondrial membrane 22 (TOM22, magenta, middle panel). Right panel displays overlay of Sirt4-sfGFP and mCherry-TOM22. Scale bar represents 2.5 µm. Squares in the upper right corner of each image show the whole cell. Scale bar represents 10 µm. Pearson correlation coefficient: 0.74 (mean) with a standard deviation of 0.10. ( b ) Representative line scan of an individual mitochondrion as demonstrated in ( a ) (left panel, white dashed line). Curves show respective fluorescence intensity of the line scan. ( c ) Identical experiments as those in ( a ), but using tetramethylrhodamine methyl ester (TMRM, magenta). Pearson correlation coefficient: 0.34 (mean) with a standard deviation of 0.14. n = 3. ( d ) Identical experiments as those in ( b ), but using TMRM (magenta). ( e ) Identical experiments as those in ( a ), but using mitochondrial matrix targeted <t>DsRed</t> <t>(mtDsRed).</t> Pearson correlation coefficient: 0.33 (mean) with a standard deviation of 0.07 (magenta). ( f ) Identical experiments as those in ( b ), using mtDsRed instead (magenta). Shown experiments are representative of 3 independent experiments, encompassing 30 different cells.
    Dsred, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    IDEX Health & Science dsred
    Sirt4-sfGFP is exclusively located at the outer mitochondrial membrane. ( a ) Representative SIM images of HeLa cells expressing Sirt4-sfGFP (green, left panel) and mCherry-translocase of the outer mitochondrial membrane 22 (TOM22, magenta, middle panel). Right panel displays overlay of Sirt4-sfGFP and mCherry-TOM22. Scale bar represents 2.5 µm. Squares in the upper right corner of each image show the whole cell. Scale bar represents 10 µm. Pearson correlation coefficient: 0.74 (mean) with a standard deviation of 0.10. ( b ) Representative line scan of an individual mitochondrion as demonstrated in ( a ) (left panel, white dashed line). Curves show respective fluorescence intensity of the line scan. ( c ) Identical experiments as those in ( a ), but using tetramethylrhodamine methyl ester (TMRM, magenta). Pearson correlation coefficient: 0.34 (mean) with a standard deviation of 0.14. n = 3. ( d ) Identical experiments as those in ( b ), but using TMRM (magenta). ( e ) Identical experiments as those in ( a ), but using mitochondrial matrix targeted <t>DsRed</t> <t>(mtDsRed).</t> Pearson correlation coefficient: 0.33 (mean) with a standard deviation of 0.07 (magenta). ( f ) Identical experiments as those in ( b ), using mtDsRed instead (magenta). Shown experiments are representative of 3 independent experiments, encompassing 30 different cells.
    Dsred, supplied by IDEX Health & Science, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Jackson Immuno dsred
    Immunofluorescence staining of differentiated human bronchial epithelial cells. Confocal microscopy immunofluorescence analysis of differentiated human bronchial epithelial cells. Figs A, B, C show XY planes, Figs D, E, F show XZ planes. (A, D) Cells were infected from the apical side with HAdV-B14p1 at a moi of 10 (TCID 50 /cell), fixated 4 days p.i. and stained for HAdV in green (FITC conjugated antibody) and the desmoglein 2 (DSG2 receptor) in red <t>(dsRed</t> antibody), the nucleus was counterstained in blue (DAPI). (B, E) Differentiated human bronchial epithelial cells stained for DSG2 receptor in red (dsRed) and occludin (OCLN) a tight junction marker in green (FITC), nucleus was counterstained in blue (DAPI). (C, F) Differentiated human bronchial epithelial cells were stained for CAR receptor in red (dsRed) and the tight junction marker OCLN in green (FITC) and the nucleus in blue (DAPI).
    Dsred, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa fp583 gene
    Subcellular localization of PTS1- and PTS2-targeted proteins in yor084w Δ and pex25 Δ cells. <t>DsRed-PTS1</t> and PTS2–GFP reporters were examined by fluorescence microscopy. DsRed-PTS1 localized to numerous small peroxisomes in pex7 Δ and yor084 Δ yeast cells. A similar localization was seen with PTS2–GFP in pex5 Δ and yor084 Δ cells. A diffuse cytoplasmic localization of these reporters was observed in strains lacking components necessary for the targeting of either PTS1 ( pex5 Δ) or PTS2 ( pex7 Δ). In pex11 Δ cells, both reporters accumulated in a few large peroxisomes as reported previously ( Erdmann and Blobel, 1995 ). In pex25 Δ cells, a heterogeneous population was observed; some cells displayed a diffuse localization of the reporters, and some cells showed accumulation of the reporters to a few large peroxisomes. Bar, 10 μm.
    Fp583 Gene, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa dsred er
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Dsred Er, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 143 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Olympus dsred filter
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Dsred Filter, supplied by Olympus, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa dsred n1
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Dsred N1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson monomeric dsred
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Monomeric Dsred, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Addgene inc pmxs dsred
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Pmxs Dsred, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson dsred protein
    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with <t>pCAG-CRE-ires-EGFP;</t> anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either <t>dsRED-Golgi</t> (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.
    Dsred Protein, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa mito dsred
    ( a ) U251 DEVD <t>Mito</t> Ds cells were grown on 96-well glass-bottom plates and treated with different drugs. High-throughput imaging for ratio and <t>Mito-DsRed</t> was carried out as described using pathway bioimager. A representative montage image of ECFP–EYFP Ratio and a merged channel of ECFP/EYFP/DsRed and scatter plot generated after proper segmentation and analysis for untreated control and cisplatin (50 μ g/ml)-treated samples is shown. ( b ) The merged images and scatter plot from high-throughput imager for cells treated with indicated drugs are shown. ( c ) U251 DEVD Mito Ds cells grown in 96-well glass-bottom plates were treated with necrosis-inducing agent, H 2 O 2 (2.5 mM). Merged image and scatter plot from high-throughput imager for treated and control cells are shown.
    Mito Dsred, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa monomeric dsred
    ( a ) U251 DEVD <t>Mito</t> Ds cells were grown on 96-well glass-bottom plates and treated with different drugs. High-throughput imaging for ratio and <t>Mito-DsRed</t> was carried out as described using pathway bioimager. A representative montage image of ECFP–EYFP Ratio and a merged channel of ECFP/EYFP/DsRed and scatter plot generated after proper segmentation and analysis for untreated control and cisplatin (50 μ g/ml)-treated samples is shown. ( b ) The merged images and scatter plot from high-throughput imager for cells treated with indicated drugs are shown. ( c ) U251 DEVD Mito Ds cells grown in 96-well glass-bottom plates were treated with necrosis-inducing agent, H 2 O 2 (2.5 mM). Merged image and scatter plot from high-throughput imager for treated and control cells are shown.
    Monomeric Dsred, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Confocal images showing localization of GFP:TGBp2 with actin, Golgi, and localization of GFP:TGBp1. A to C, Cells expressing GFP:TGBp2 and DsRed:Talin. Arrows point to vesicles along actin filaments. D to F, Cells expressing GFP:TGBp2 and GFP:Talin. Yellow arrowheads point to vesicles. F, Vesicles in cells treated with latrunculin B. G to J, Tobacco epidermal cells cobombarded with GFP-TGBp2 and DsRed-ST. Note that the GFP-TGBp2 vesicles (arrowhead) do not colocalize with the DsRed-ST decorated Golgi (arrows). K, Cells expressing GFP:TGBp2. L, Cells expressing GFP:TGBp1. Bars represent 10 μ m.

    Journal: Plant Physiology

    Article Title: The Potato Virus X TGBp2 Movement Protein Associates with Endoplasmic Reticulum-Derived Vesicles during Virus Infection 1

    doi: 10.1104/pp.105.066019

    Figure Lengend Snippet: Confocal images showing localization of GFP:TGBp2 with actin, Golgi, and localization of GFP:TGBp1. A to C, Cells expressing GFP:TGBp2 and DsRed:Talin. Arrows point to vesicles along actin filaments. D to F, Cells expressing GFP:TGBp2 and GFP:Talin. Yellow arrowheads point to vesicles. F, Vesicles in cells treated with latrunculin B. G to J, Tobacco epidermal cells cobombarded with GFP-TGBp2 and DsRed-ST. Note that the GFP-TGBp2 vesicles (arrowhead) do not colocalize with the DsRed-ST decorated Golgi (arrows). K, Cells expressing GFP:TGBp2. L, Cells expressing GFP:TGBp1. Bars represent 10 μ m.

    Article Snippet: The pBIN-mGFP5-ER plasmid was obtained from Dr. J. Hasselof (Medical Research Council Laboratory of Molecular Biology, Cambridge, UK; ); pDsRed-ST was a generous gift from Dr. R. Cyr (Pennsylvania State University, University Park, PA; ). pRTL2-GFP:Talin and -DsRed:Talin contain either enhanced GFP or DsRed (living colors; CLONTECH Laboratories, Palo Alto, CA) fused with the coding sequence of the F-actin-binding domain of mouse Talin ( ; ).

    Techniques: Expressing

    The distribution of Golgi-DsRed (A and B) and spGFP-VSR3 (C and D) in the wild type and glup4 (EM425). A and C, The wild type (WT). B and D, glup4 (EM425). Transgenic wild-type and glup4 lines expressing Golgi-DsRed (β-galactosyltransferase) or

    Journal: Plant Physiology

    Article Title: The Small GTPase Rab5a Is Essential for Intracellular Transport of Proglutelin from the Golgi Apparatus to the Protein Storage Vacuole and Endosomal Membrane Organization in Developing Rice Endosperm 1The Small GTPase Rab5a Is Essential for Intracellular Transport of Proglutelin from the Golgi Apparatus to the Protein Storage Vacuole and Endosomal Membrane Organization in Developing Rice Endosperm 1 [C]The Small GTPase Rab5a Is Essential for Intracellular Transport of Proglutelin from the Golgi Apparatus to the Protein Storage Vacuole and Endosomal Membrane Organization in Developing Rice Endosperm 1 [C] [W]The Small GTPase Rab5a Is Essential for Intracellular Transport of Proglutelin from the Golgi Apparatus to the Protein Storage Vacuole and Endosomal Membrane Organization in Developing Rice Endosperm 1 [C] [W] [OA]

    doi: 10.1104/pp.111.180505

    Figure Lengend Snippet: The distribution of Golgi-DsRed (A and B) and spGFP-VSR3 (C and D) in the wild type and glup4 (EM425). A and C, The wild type (WT). B and D, glup4 (EM425). Transgenic wild-type and glup4 lines expressing Golgi-DsRed (β-galactosyltransferase) or

    Article Snippet: The GFP of the fusion protein was replaced with DsRed-Monomer (Clontech), generating Golgi-DsRed.

    Techniques: Transgenic Assay, Expressing

    Development and characterization of QuasAr3. (a) Screening pipeline. Rationally designed constructs were cloned in the Optopatch configuration, expressed in primary neurons and tested for spike SNR using light-induced spikes. Constructs with improved SNR were then expressed in vivo using in utero electroporation (IUE) and tested for spike SNR in acute slices. The process was repeated iteratively. (b) Examples of SNR measurements in cultured neurons. Left: wide-field epifluorescence images of GFP fused to CheRiff, an opsin with excellent membrane trafficking. Middle: Fluorescence of QuasAr mutants. Scale bar 10 μm. Right: QuasAr fluorescence transients in response to optogenetically induced spikes (10 ms blue light stimulation at 1 mW/mm 2 ). Each construct was tested on at least 5 cultured neurons. (c) Hierarchical screen for improved membrane trafficking of QuasAr variants (see Methods for details). Diagram: schematic of the FCK_DuEx1.0 construct and overview of the screening pipeline. E. coli colonies were transformed with libraries in FCK_DuEx1.0. The colonies with the brightest fluorescence were picked for lentivirus production and secondary screening in primary neuronal culture. Images: example images of the FP channel of QuasAr2-FP fusions: i. mOrange; ii. mRuby2; iii. mKate2; iv. Citrine. Scale bar 10 μm. (d) SNR of N-terminal modifications compared with QuasAr2. All constructs showed reduced SNR (see Methods for details). (e) Replacing mOrange2 with Citrine as a fusion protein improved the trafficking only with two specific linkers. (f) Adding additional TS sequences at the linker and C terminal improved the spike SNR. (g) The mutation K171R increased the QuasAr expression level, quantified by normalizing QuasAr fluorescence by the fluorescence of the co-expressed CheRiff-GFP. (d-g) all error bars are mean ± s.e.m., 1-tail t -test. (h) Top: diagram of the QuasAr2 and QuasAr3 constructs. Bottom: Confocal images of brain slices expressing QuasAr2 and QuasAr3. Scale bar 500 μm. Insets: single cell bodies, scale bar 10 μm. Representative images from N = 2 mice (QuasAr2) and N = 3 mice (QuasAr3). (i) Confocal images of brain slices expressing Cre-dependent QuasAr3 with sparsity controlled by co-expression of hSyn-Cre. Scale bar 50 μm. (j) Simultaneous fluorescence and patch clamp recordings from two neurons expressing QuasAr3 using AAV virus in acute brain slice. Left: image of QuasAr3 fluorescence in the soma. Scale bar 10 μm. Middle: spiking during ramp current injection. Right: mean spike, overlay of fluorescence and voltage. Inset: boxed regions showing correspondence of optical and electrical recordings of sub-threshold voltage overlaid. See Extended Data Fig. 3 for statistics. Scale bar 10 μm.

    Journal: Nature

    Article Title: Voltage imaging and optogenetics reveal behavior dependent changes in hippocampal dynamics

    doi: 10.1038/s41586-019-1166-7

    Figure Lengend Snippet: Development and characterization of QuasAr3. (a) Screening pipeline. Rationally designed constructs were cloned in the Optopatch configuration, expressed in primary neurons and tested for spike SNR using light-induced spikes. Constructs with improved SNR were then expressed in vivo using in utero electroporation (IUE) and tested for spike SNR in acute slices. The process was repeated iteratively. (b) Examples of SNR measurements in cultured neurons. Left: wide-field epifluorescence images of GFP fused to CheRiff, an opsin with excellent membrane trafficking. Middle: Fluorescence of QuasAr mutants. Scale bar 10 μm. Right: QuasAr fluorescence transients in response to optogenetically induced spikes (10 ms blue light stimulation at 1 mW/mm 2 ). Each construct was tested on at least 5 cultured neurons. (c) Hierarchical screen for improved membrane trafficking of QuasAr variants (see Methods for details). Diagram: schematic of the FCK_DuEx1.0 construct and overview of the screening pipeline. E. coli colonies were transformed with libraries in FCK_DuEx1.0. The colonies with the brightest fluorescence were picked for lentivirus production and secondary screening in primary neuronal culture. Images: example images of the FP channel of QuasAr2-FP fusions: i. mOrange; ii. mRuby2; iii. mKate2; iv. Citrine. Scale bar 10 μm. (d) SNR of N-terminal modifications compared with QuasAr2. All constructs showed reduced SNR (see Methods for details). (e) Replacing mOrange2 with Citrine as a fusion protein improved the trafficking only with two specific linkers. (f) Adding additional TS sequences at the linker and C terminal improved the spike SNR. (g) The mutation K171R increased the QuasAr expression level, quantified by normalizing QuasAr fluorescence by the fluorescence of the co-expressed CheRiff-GFP. (d-g) all error bars are mean ± s.e.m., 1-tail t -test. (h) Top: diagram of the QuasAr2 and QuasAr3 constructs. Bottom: Confocal images of brain slices expressing QuasAr2 and QuasAr3. Scale bar 500 μm. Insets: single cell bodies, scale bar 10 μm. Representative images from N = 2 mice (QuasAr2) and N = 3 mice (QuasAr3). (i) Confocal images of brain slices expressing Cre-dependent QuasAr3 with sparsity controlled by co-expression of hSyn-Cre. Scale bar 50 μm. (j) Simultaneous fluorescence and patch clamp recordings from two neurons expressing QuasAr3 using AAV virus in acute brain slice. Left: image of QuasAr3 fluorescence in the soma. Scale bar 10 μm. Middle: spiking during ramp current injection. Right: mean spike, overlay of fluorescence and voltage. Inset: boxed regions showing correspondence of optical and electrical recordings of sub-threshold voltage overlaid. See Extended Data Fig. 3 for statistics. Scale bar 10 μm.

    Article Snippet: All AAV plasmids were deposited with Addgene as follows: hSyn-QuasAr2-mOrange ( , Addgene #107705) hSyn-QuasAr3-Citrine-P2A-CheRiff ( , Addgene #107700) CAG-FLEX-QuasAr3-Citrine ( - , , Addgene #107701) CAG-FLEX-paQuasAr3-Citrine ( - , , Addgene #107702) CAG-FLEX-paQuasAr3s-Citrine ( - , , , - , Addgene #107703) hSyn-Dio-paQuasAr3s-Citrine-P2A-CheRiff-s ( , , , Addgene #107704)

    Techniques: Construct, Clone Assay, In Vivo, In Utero, Electroporation, Cell Culture, Fluorescence, Mass Spectrometry, Transformation Assay, Mutagenesis, Expressing, Mouse Assay, Patch Clamp, Slice Preparation, Injection

    DsRed fluorescence imaging (A) shows the location of 4T1.2neu/R primary tumor in Balb/c mice, 2 weeks after subcutaneous inoculation into the mammary pad. NIR fluorescence (B) and PET/CT (C) imaging performed 24 hours after intravenous administration

    Journal: Translational Oncology

    Article Title: Detection of Cancer Metastases with a Dual-labeled Near-Infrared/Positron Emission Tomography Imaging Agent 1Detection of Cancer Metastases with a Dual-labeled Near-Infrared/Positron Emission Tomography Imaging Agent 1 2

    doi:

    Figure Lengend Snippet: DsRed fluorescence imaging (A) shows the location of 4T1.2neu/R primary tumor in Balb/c mice, 2 weeks after subcutaneous inoculation into the mammary pad. NIR fluorescence (B) and PET/CT (C) imaging performed 24 hours after intravenous administration

    Article Snippet: The fluorescence was collected through holographic (model HNPF-785.0-2.0 for NIR and model 568.2–6 for DsRed; Kaiser Optical Systems, Inc, Ann Arbor, MI) and interference (model 830.0-2.0 for NIR, Image Quality [Andover Corp, Salem, NH] and model HQ610/60M for DsRed [Chroma Technology Corp, Bellows Falls, VT]) filters placed before a Nikon camera lens (Nikkor 28 mm; Nikon, Tokyo, Japan).

    Techniques: Fluorescence, Imaging, Mouse Assay, Positron Emission Tomography

    Fig. 4. In vivo localization of Hmo1–GFP and Fob1–GFP fusion proteins. ( A ) were grown at 30°C to mid-log phase (OD 600 = 0.5) in YPD. Cells were washed with water. Nucleoplasm and nucleolus were visualized using DNA staining and Nop1 staining, respectively. GFP (green), DsRed (red) and Hoechst 33352 (blue) signals were monitored by fluorescence microscopy as described in Materials and methods. Cells were examined by Nomarski imaging. ( B ) Fob1 and Hmo1 co-localize. Fob1 and Hmo1 were tagged with different GFP spectral variants, CFP and YFP, and visualized in a diploid strain prepared by crossing strains BMA-Fob1–CFP and BMA-HMO1–YFP as described in Materials and methods (scale bars correspond to 2 µm).

    Journal: The EMBO Journal

    Article Title: Hmo1, an HMG-box protein, belongs to the yeast ribosomal DNA transcription system

    doi: 10.1093/emboj/cdf539

    Figure Lengend Snippet: Fig. 4. In vivo localization of Hmo1–GFP and Fob1–GFP fusion proteins. ( A ) were grown at 30°C to mid-log phase (OD 600 = 0.5) in YPD. Cells were washed with water. Nucleoplasm and nucleolus were visualized using DNA staining and Nop1 staining, respectively. GFP (green), DsRed (red) and Hoechst 33352 (blue) signals were monitored by fluorescence microscopy as described in Materials and methods. Cells were examined by Nomarski imaging. ( B ) Fob1 and Hmo1 co-localize. Fob1 and Hmo1 were tagged with different GFP spectral variants, CFP and YFP, and visualized in a diploid strain prepared by crossing strains BMA-Fob1–CFP and BMA-HMO1–YFP as described in Materials and methods (scale bars correspond to 2 µm).

    Article Snippet: Fluorescent signals were collected with single band pass filters for excitation of DsRed (XF137-2, Omega optical), GFP (GFP, Leica), YFP (XF104, Omega Optical), CFP (XF114-2, Omega optical) and Hoechst 33352 (A, Leica).

    Techniques: In Vivo, Staining, Fluorescence, Microscopy, Imaging

    125–267, which contains TMD4R but lacks TMD2R, only localizes to peroxisomes in uninduced cells. Colocalization of Pmp47-GFP fusion proteins and DsRed-AKL is shown. (A) Representative single cells from glucose and glycerol cultures. (B) Representative cells from oleate cultures.

    Journal: Molecular Biology of the Cell

    Article Title: Multiple Targeting Modules on Peroxisomal Proteins Are Not Redundant: Discrete Functions of Targeting Signals within Pmp47 and Pex8p

    doi: 10.1091/mbc.E03-11-0810

    Figure Lengend Snippet: 125–267, which contains TMD4R but lacks TMD2R, only localizes to peroxisomes in uninduced cells. Colocalization of Pmp47-GFP fusion proteins and DsRed-AKL is shown. (A) Representative single cells from glucose and glycerol cultures. (B) Representative cells from oleate cultures.

    Article Snippet: Green fluorescent protein (GFP) was visualized with fluorescein isothiocyanate filter set, and DsRed (BD Biosciences Clontech, Palo Alto, CA) and MitoTracker Red CMXRos (Molecular Probes, Eugene, OR) were observed with the Texas Red filter set.

    Techniques:

    Effect of TIGS of candidate genes on the number of DsRed fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and GFP expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P

    Journal: Journal of Experimental Botany

    Article Title: A transient assay system for the assessment of cell-autonomous gene function in dehydration-stressed barley

    doi: 10.1093/jxb/ern186

    Figure Lengend Snippet: Effect of TIGS of candidate genes on the number of DsRed fluorescent cells in non-stressed control and dehydration-stressed leaf segments. Leaf segments were co-bombarded with DsRed and GFP expression plasmids, plus an RNAi construct targeting the barley genes listed below the graph. If no annotated target gene is known in barley, the name of the closest homologue from another plant species is given (see also Table 2 ). After a dehydration stress period of 4 d, the number of DsRed fluorescent cells was counted and normalized to GFP. During the stress period, control leaves were incubated on water–agar. (A) Number of DsRed fluorescent cells in non-stressed control leaves in the presence of the empty RNAi vector pIPKTA30 or the RNAi test construct. Mean ±SEM of five independent experiments, except for RNAi constructs targeting HvDRF1 , HvDhn6 , HVA1 , and HvNHX1 where combined data from two series of five experiments each are shown. An asterisk indicates a statistically significant difference ( P

    Article Snippet: Fluorescence microscopy of GFP and DsRED For GFP and DsRed detection, single-cell fluorescence was examined 24 h (GFP) or 120 h (DsRed) post-bombardment by using a Zeiss Axioplan 2 imaging microscope with the filter set BP450–490 excitation, FT510 beam splitter, BP515–565 emission for GFP fluorescence, and the filter set BP546/12 excitation, FT580 beam splitter, BP590 emission for DsRed detection.

    Techniques: Expressing, Construct, Incubation, Plasmid Preparation

    Kinetics of transient GFP and DsRed expression, reflected by the number of fluorescing epidermal cells per bombardment. The same bombarded leaf segments were repeatedly counted for the number of green- or red-fluorescing epidermal cells at the times indicated. Mean ±SEM of five leaf segments. The same experiment was repeated with similar results.

    Journal: Journal of Experimental Botany

    Article Title: A transient assay system for the assessment of cell-autonomous gene function in dehydration-stressed barley

    doi: 10.1093/jxb/ern186

    Figure Lengend Snippet: Kinetics of transient GFP and DsRed expression, reflected by the number of fluorescing epidermal cells per bombardment. The same bombarded leaf segments were repeatedly counted for the number of green- or red-fluorescing epidermal cells at the times indicated. Mean ±SEM of five leaf segments. The same experiment was repeated with similar results.

    Article Snippet: Fluorescence microscopy of GFP and DsRED For GFP and DsRed detection, single-cell fluorescence was examined 24 h (GFP) or 120 h (DsRed) post-bombardment by using a Zeiss Axioplan 2 imaging microscope with the filter set BP450–490 excitation, FT510 beam splitter, BP515–565 emission for GFP fluorescence, and the filter set BP546/12 excitation, FT580 beam splitter, BP590 emission for DsRed detection.

    Techniques: Expressing

    Wild-type mouse HSPC-derived cells deliver frataxin and Cox8 to FRDA cells in vitro and in vivo ( A and B ). Scale bars, 10 μm. ( C ) Representative confocal images of brain sections from a YG8R mouse transplanted with DsRed + ) and those of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + /Cox8-GFP + for 3D visualization). Scale bars, 10 μm. ( D ) Representative confocal images of a spinal cord section from a YG8R mouse transplanted with DsRed + /Cox8-GFP + HSPCs at 7 months after transplantation, labeled with an anti-NeuN antibody (blue). White arrows depict Cox8-GFP within branch extensions of the DsRed + microglial cell. Scale bar, 5 μm. ( E ) Quantification of neurons containing Cox8-GFP in the cervical spinal cord gray matter of YG8R mice transplanted with DsRed + /Cox8-GFP + for the description of the automatic unbiased quantification method). ( F ) Representative confocal images of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + HSPCs transduced with LV-hFXN-GFP at 7 months after transplantation and stained with anti-mCherry (red) and anti-NeuN (blue) antibodies. Scale bars, 10 μm.

    Journal: Science translational medicine

    Article Title: Transplantation of wild-type mouse hematopoietic stem and progenitor cells ameliorates deficits in a mouse model of Friedreich’s ataxia

    doi: 10.1126/scitranslmed.aaj2347

    Figure Lengend Snippet: Wild-type mouse HSPC-derived cells deliver frataxin and Cox8 to FRDA cells in vitro and in vivo ( A and B ). Scale bars, 10 μm. ( C ) Representative confocal images of brain sections from a YG8R mouse transplanted with DsRed + ) and those of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + /Cox8-GFP + for 3D visualization). Scale bars, 10 μm. ( D ) Representative confocal images of a spinal cord section from a YG8R mouse transplanted with DsRed + /Cox8-GFP + HSPCs at 7 months after transplantation, labeled with an anti-NeuN antibody (blue). White arrows depict Cox8-GFP within branch extensions of the DsRed + microglial cell. Scale bar, 5 μm. ( E ) Quantification of neurons containing Cox8-GFP in the cervical spinal cord gray matter of YG8R mice transplanted with DsRed + /Cox8-GFP + for the description of the automatic unbiased quantification method). ( F ) Representative confocal images of brain and spinal cord sections from a YG8R mouse transplanted with DsRed + HSPCs transduced with LV-hFXN-GFP at 7 months after transplantation and stained with anti-mCherry (red) and anti-NeuN (blue) antibodies. Scale bars, 10 μm.

    Article Snippet: Transgenic mice constitutively expressing GFP [C57BL/6-Tg(ACTB-EGFP)1Osb/J] or DsRed [B6.Cg-Tg(CAG-DsRed*MST) 1Nagy/J] were also purchased from the Jackson Laboratory.

    Techniques: Derivative Assay, In Vitro, In Vivo, Transplantation Assay, Labeling, Mouse Assay, Transduction, Staining

    Sirt4-sfGFP is exclusively located at the outer mitochondrial membrane. ( a ) Representative SIM images of HeLa cells expressing Sirt4-sfGFP (green, left panel) and mCherry-translocase of the outer mitochondrial membrane 22 (TOM22, magenta, middle panel). Right panel displays overlay of Sirt4-sfGFP and mCherry-TOM22. Scale bar represents 2.5 µm. Squares in the upper right corner of each image show the whole cell. Scale bar represents 10 µm. Pearson correlation coefficient: 0.74 (mean) with a standard deviation of 0.10. ( b ) Representative line scan of an individual mitochondrion as demonstrated in ( a ) (left panel, white dashed line). Curves show respective fluorescence intensity of the line scan. ( c ) Identical experiments as those in ( a ), but using tetramethylrhodamine methyl ester (TMRM, magenta). Pearson correlation coefficient: 0.34 (mean) with a standard deviation of 0.14. n = 3. ( d ) Identical experiments as those in ( b ), but using TMRM (magenta). ( e ) Identical experiments as those in ( a ), but using mitochondrial matrix targeted DsRed (mtDsRed). Pearson correlation coefficient: 0.33 (mean) with a standard deviation of 0.07 (magenta). ( f ) Identical experiments as those in ( b ), using mtDsRed instead (magenta). Shown experiments are representative of 3 independent experiments, encompassing 30 different cells.

    Journal: Cells

    Article Title: Visualization of Sirtuin 4 Distribution between Mitochondria and the Nucleus, Based on Bimolecular Fluorescence Self-Complementation

    doi: 10.3390/cells8121583

    Figure Lengend Snippet: Sirt4-sfGFP is exclusively located at the outer mitochondrial membrane. ( a ) Representative SIM images of HeLa cells expressing Sirt4-sfGFP (green, left panel) and mCherry-translocase of the outer mitochondrial membrane 22 (TOM22, magenta, middle panel). Right panel displays overlay of Sirt4-sfGFP and mCherry-TOM22. Scale bar represents 2.5 µm. Squares in the upper right corner of each image show the whole cell. Scale bar represents 10 µm. Pearson correlation coefficient: 0.74 (mean) with a standard deviation of 0.10. ( b ) Representative line scan of an individual mitochondrion as demonstrated in ( a ) (left panel, white dashed line). Curves show respective fluorescence intensity of the line scan. ( c ) Identical experiments as those in ( a ), but using tetramethylrhodamine methyl ester (TMRM, magenta). Pearson correlation coefficient: 0.34 (mean) with a standard deviation of 0.14. n = 3. ( d ) Identical experiments as those in ( b ), but using TMRM (magenta). ( e ) Identical experiments as those in ( a ), but using mitochondrial matrix targeted DsRed (mtDsRed). Pearson correlation coefficient: 0.33 (mean) with a standard deviation of 0.07 (magenta). ( f ) Identical experiments as those in ( b ), using mtDsRed instead (magenta). Shown experiments are representative of 3 independent experiments, encompassing 30 different cells.

    Article Snippet: For the cloning of mtDsRed, DsRed (plasmid number 11151) was purchased from Addgene and isolated via PCR-amplification.

    Techniques: Expressing, Standard Deviation, Fluorescence

    Immunofluorescence staining of differentiated human bronchial epithelial cells. Confocal microscopy immunofluorescence analysis of differentiated human bronchial epithelial cells. Figs A, B, C show XY planes, Figs D, E, F show XZ planes. (A, D) Cells were infected from the apical side with HAdV-B14p1 at a moi of 10 (TCID 50 /cell), fixated 4 days p.i. and stained for HAdV in green (FITC conjugated antibody) and the desmoglein 2 (DSG2 receptor) in red (dsRed antibody), the nucleus was counterstained in blue (DAPI). (B, E) Differentiated human bronchial epithelial cells stained for DSG2 receptor in red (dsRed) and occludin (OCLN) a tight junction marker in green (FITC), nucleus was counterstained in blue (DAPI). (C, F) Differentiated human bronchial epithelial cells were stained for CAR receptor in red (dsRed) and the tight junction marker OCLN in green (FITC) and the nucleus in blue (DAPI).

    Journal: PLoS ONE

    Article Title: Effective Apical Infection of Differentiated Human Bronchial Epithelial Cells and Induction of Proinflammatory Chemokines by the Highly Pneumotropic Human Adenovirus Type 14p1

    doi: 10.1371/journal.pone.0131201

    Figure Lengend Snippet: Immunofluorescence staining of differentiated human bronchial epithelial cells. Confocal microscopy immunofluorescence analysis of differentiated human bronchial epithelial cells. Figs A, B, C show XY planes, Figs D, E, F show XZ planes. (A, D) Cells were infected from the apical side with HAdV-B14p1 at a moi of 10 (TCID 50 /cell), fixated 4 days p.i. and stained for HAdV in green (FITC conjugated antibody) and the desmoglein 2 (DSG2 receptor) in red (dsRed antibody), the nucleus was counterstained in blue (DAPI). (B, E) Differentiated human bronchial epithelial cells stained for DSG2 receptor in red (dsRed) and occludin (OCLN) a tight junction marker in green (FITC), nucleus was counterstained in blue (DAPI). (C, F) Differentiated human bronchial epithelial cells were stained for CAR receptor in red (dsRed) and the tight junction marker OCLN in green (FITC) and the nucleus in blue (DAPI).

    Article Snippet: Secondary antibodies were anti rabbit conjugated with FITC and anti mouse conjugated with dsRed (1:200 diluted, Jackson, Immunoresearch, West Grove, PA).

    Techniques: Immunofluorescence, Staining, Confocal Microscopy, Infection, Marker

    Subcellular localization of PTS1- and PTS2-targeted proteins in yor084w Δ and pex25 Δ cells. DsRed-PTS1 and PTS2–GFP reporters were examined by fluorescence microscopy. DsRed-PTS1 localized to numerous small peroxisomes in pex7 Δ and yor084 Δ yeast cells. A similar localization was seen with PTS2–GFP in pex5 Δ and yor084 Δ cells. A diffuse cytoplasmic localization of these reporters was observed in strains lacking components necessary for the targeting of either PTS1 ( pex5 Δ) or PTS2 ( pex7 Δ). In pex11 Δ cells, both reporters accumulated in a few large peroxisomes as reported previously ( Erdmann and Blobel, 1995 ). In pex25 Δ cells, a heterogeneous population was observed; some cells displayed a diffuse localization of the reporters, and some cells showed accumulation of the reporters to a few large peroxisomes. Bar, 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: Transcriptome profiling to identify genes involved in peroxisome assembly and function

    doi: 10.1083/jcb.200204059

    Figure Lengend Snippet: Subcellular localization of PTS1- and PTS2-targeted proteins in yor084w Δ and pex25 Δ cells. DsRed-PTS1 and PTS2–GFP reporters were examined by fluorescence microscopy. DsRed-PTS1 localized to numerous small peroxisomes in pex7 Δ and yor084 Δ yeast cells. A similar localization was seen with PTS2–GFP in pex5 Δ and yor084 Δ cells. A diffuse cytoplasmic localization of these reporters was observed in strains lacking components necessary for the targeting of either PTS1 ( pex5 Δ) or PTS2 ( pex7 Δ). In pex11 Δ cells, both reporters accumulated in a few large peroxisomes as reported previously ( Erdmann and Blobel, 1995 ). In pex25 Δ cells, a heterogeneous population was observed; some cells displayed a diffuse localization of the reporters, and some cells showed accumulation of the reporters to a few large peroxisomes. Bar, 10 μm.

    Article Snippet: Plasmids The following plasmids were described previously: pRS315 CEN/LEU2 and pRS316 CEN/URA3 , pProtA/HIS5 , pBluescript II SK (Stratagene), and pGFP/HIS5 ( ). pDsRed-PTS1 was constructed by PCR directed mutagenesis ( ) to yield the Discosoma sp. FP583 gene (DsRed; CLONETECH Laboratories, Inc.), containing a 3′ extension encoding a tripeptide PTS1 (Ser-Lys-Leu) flanked by the ORE-containing FAA2 promoter (−1 to −398) and terminator (+2236 to +2690).

    Techniques: Fluorescence, Microscopy

    Cellular distributions of Pex25p and Yor084p. ( A ) The DsRed-PTS1 reporter colocalizes with Fox2–GFP chimera to peroxisomes. (B) The subcellular distributions of protein A chimeras were compared with that of DsRed-PTS1 in oleate-induced haploid cells (Pex11-pA and Pex25-pA) and heterozygous diploid cells (Yor084-pA) by double-labeling immunofluorescence microscopy. Bar, 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: Transcriptome profiling to identify genes involved in peroxisome assembly and function

    doi: 10.1083/jcb.200204059

    Figure Lengend Snippet: Cellular distributions of Pex25p and Yor084p. ( A ) The DsRed-PTS1 reporter colocalizes with Fox2–GFP chimera to peroxisomes. (B) The subcellular distributions of protein A chimeras were compared with that of DsRed-PTS1 in oleate-induced haploid cells (Pex11-pA and Pex25-pA) and heterozygous diploid cells (Yor084-pA) by double-labeling immunofluorescence microscopy. Bar, 10 μm.

    Article Snippet: Plasmids The following plasmids were described previously: pRS315 CEN/LEU2 and pRS316 CEN/URA3 , pProtA/HIS5 , pBluescript II SK (Stratagene), and pGFP/HIS5 ( ). pDsRed-PTS1 was constructed by PCR directed mutagenesis ( ) to yield the Discosoma sp. FP583 gene (DsRed; CLONETECH Laboratories, Inc.), containing a 3′ extension encoding a tripeptide PTS1 (Ser-Lys-Leu) flanked by the ORE-containing FAA2 promoter (−1 to −398) and terminator (+2236 to +2690).

    Techniques: Labeling, Immunofluorescence, Microscopy

    Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with pCAG-CRE-ires-EGFP; anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either dsRED-Golgi (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.

    Journal: PLoS ONE

    Article Title: Selective Gene Expression by Postnatal Electroporation during Olfactory Interneuron Neurogenesis

    doi: 10.1371/journal.pone.0001517

    Figure Lengend Snippet: Electroporation is effective for the expression of multiple constructs, in transgenic animals, and in rats. A. 20 µm section of an olfactory bulb of a P6 mouse 5 days post-electroporation with pCAG-CRE-ires-EGFP; anti-CRE staining labels the nucleus (red) and GFP (green) labels the cytoplasm with a 10X magnified view of the cell body. Scale = 25 µm. B–C. Z projections of 60 µm sections from olfactory bulbs co-electroporated with GFP and either dsRED-Golgi (B; 5 days post-electroporation) or dsRED-ER (C; 14 days post electroporation) showing labeled organelles. 10X magnified views of highlighted regions show perinuclear golgi staining in the cell body and punctate ER staining at the base of dendritic spines. Scale = 25 µm. D–E. Electroporation of OMP-GFP mice with RFP plasmid. Z-projections of 60 µm sections showing the morphology of single periglomerular cells (red) and the glomeruli they innervate (green). 2 types of PG cells are shown, one that innervates 3 glomeruli (E; scale = 25 µm) and another other with its processes are confined to a single glomerulus (F; scale = 50 µm). F. Highly efficient electroporation of rats. Z-projection of 60 µm section from a P27 rat olfactory bulb electroporated at P1 showing widespread ectopic expression in all layers of the olfactory bulb. TOTO3 (blue) labels all nuclei. Scale = 50 µm.

    Article Snippet: A BglII-GCamp2-NotI fragment was subcloned into pENTR1a (Invitrogen) to create pENTR1a-GCamp2, and was subsequnetly recombined with pCAG-Gt to create pCAG-GCamp2. pCAG-Gt-IRES-EGFP was created by subcloning IRES-EGFPfor pIRES-EGFP2 (Clontech) behind the reading frame cassette. nlsCRE was kindly supplied by Andreas Walz (Rockefeller University), subcloned into pENTR1a and recombined into pCAG-Gt-IRES-EGFP. dsRED-Golgi and dsRED-ER (Clonetech) were subcloned into pENTR1a and recombined into pCAG-Gt.

    Techniques: Electroporation, Expressing, Construct, Transgenic Assay, Staining, Labeling, Mouse Assay, Plasmid Preparation

    ( a ) U251 DEVD Mito Ds cells were grown on 96-well glass-bottom plates and treated with different drugs. High-throughput imaging for ratio and Mito-DsRed was carried out as described using pathway bioimager. A representative montage image of ECFP–EYFP Ratio and a merged channel of ECFP/EYFP/DsRed and scatter plot generated after proper segmentation and analysis for untreated control and cisplatin (50 μ g/ml)-treated samples is shown. ( b ) The merged images and scatter plot from high-throughput imager for cells treated with indicated drugs are shown. ( c ) U251 DEVD Mito Ds cells grown in 96-well glass-bottom plates were treated with necrosis-inducing agent, H 2 O 2 (2.5 mM). Merged image and scatter plot from high-throughput imager for treated and control cells are shown.

    Journal: Cell Death Discovery

    Article Title: A quantitative real-time approach for discriminating apoptosis and necrosis

    doi: 10.1038/cddiscovery.2016.101

    Figure Lengend Snippet: ( a ) U251 DEVD Mito Ds cells were grown on 96-well glass-bottom plates and treated with different drugs. High-throughput imaging for ratio and Mito-DsRed was carried out as described using pathway bioimager. A representative montage image of ECFP–EYFP Ratio and a merged channel of ECFP/EYFP/DsRed and scatter plot generated after proper segmentation and analysis for untreated control and cisplatin (50 μ g/ml)-treated samples is shown. ( b ) The merged images and scatter plot from high-throughput imager for cells treated with indicated drugs are shown. ( c ) U251 DEVD Mito Ds cells grown in 96-well glass-bottom plates were treated with necrosis-inducing agent, H 2 O 2 (2.5 mM). Merged image and scatter plot from high-throughput imager for treated and control cells are shown.

    Article Snippet: Expression vector for Mito-DsRed (#6975-1) was purchased from Clontech Laboratories Inc, Mountain View, CA, USA.

    Techniques: High Throughput Screening Assay, Imaging, Generated

    ( a ) U251 DEVD Mito Ds cells were exposed to different drugs for 24 h. The cells were analyzed using flow cytometer for green fluorescence at 488 nm and ratio mode for FRET at 405 nm. The necrotic cells are gated based on Mito-DsRed fluorescence against green channel. The cell with increase in ratio represents cells with caspase activation. ( b ) The percentage of cells with apoptosis and necrosis were scored from the three different flow cytometry data for the indicated drugs. Results represented are mean±S.D. ( n =3). ( c ) U251 DEVD Mito Ds cells were exposed to different drugs for 24 h. The cells were trypsinized and stained with Alexa Fluor 647 conjugated Annexin-V as described. The cells were analyzed for ratio change and annexin-V staining. The scatter plot of annexin-V versus ratio of the cells treated with indicated drugs are shown.

    Journal: Cell Death Discovery

    Article Title: A quantitative real-time approach for discriminating apoptosis and necrosis

    doi: 10.1038/cddiscovery.2016.101

    Figure Lengend Snippet: ( a ) U251 DEVD Mito Ds cells were exposed to different drugs for 24 h. The cells were analyzed using flow cytometer for green fluorescence at 488 nm and ratio mode for FRET at 405 nm. The necrotic cells are gated based on Mito-DsRed fluorescence against green channel. The cell with increase in ratio represents cells with caspase activation. ( b ) The percentage of cells with apoptosis and necrosis were scored from the three different flow cytometry data for the indicated drugs. Results represented are mean±S.D. ( n =3). ( c ) U251 DEVD Mito Ds cells were exposed to different drugs for 24 h. The cells were trypsinized and stained with Alexa Fluor 647 conjugated Annexin-V as described. The cells were analyzed for ratio change and annexin-V staining. The scatter plot of annexin-V versus ratio of the cells treated with indicated drugs are shown.

    Article Snippet: Expression vector for Mito-DsRed (#6975-1) was purchased from Clontech Laboratories Inc, Mountain View, CA, USA.

    Techniques: Flow Cytometry, Cytometry, Fluorescence, Activation Assay, Staining

    ( a ) U251 caspase sensor cell line was transfected with DsRed targeted at mitochondria to generate stable cells expressing both the probe, U251 DEVD Mito Ds cells. Representative images in brightfield, ECFP, EYFP, DsRed channels and ratio mode of the stable cells are shown. ( b ) U251 DEVD Mito Ds cells were treated with caspase activating compound doxorubicin (200 ng/ml). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using fluorescence microscope at an interval of 10 min as described in the 'Materials and Methods'. The snapshot of merged channels of ECFP/EYFP/DsRed at 3 h, 6 h, 12 h and 24 h of treatment is shown. The caspase activation is reflected from the FRET loss and subsequent increase in blue channel fluorescence. ( c ) Images representing three distinct cell populations identified after doxorubicin (200 ng/ml) treatment of U251 DEVD Mito-DsRed cell: apoptotic cells with ECFP/EYFP ratio change and retaining red fluorescence, necrotic cells without any ECFP/EYFP fluorescence and retaining red fluorescence and live cells with intact ECFP/EYFP probe without ratio change and retaining red fluorescence. In the image, 'A' represents apoptotic cells, 'N' represents necrotic cells and 'L' represents the live intact cells. ( d ) U251 DEVD Mito Ds cells were either untreated or treated with caspase activating compound for 24 h. The cells were imaged with fluorescence microscope for ratio imaging and DsRed channel. The cells with caspase activation and necrosis were identified on the basis of ratio change and DsRed fluorescence as described. The quantitative necrosis and apoptosis from three different independent experiments are shown in the graph. The results represented are mean±S.D. ( n =3).

    Journal: Cell Death Discovery

    Article Title: A quantitative real-time approach for discriminating apoptosis and necrosis

    doi: 10.1038/cddiscovery.2016.101

    Figure Lengend Snippet: ( a ) U251 caspase sensor cell line was transfected with DsRed targeted at mitochondria to generate stable cells expressing both the probe, U251 DEVD Mito Ds cells. Representative images in brightfield, ECFP, EYFP, DsRed channels and ratio mode of the stable cells are shown. ( b ) U251 DEVD Mito Ds cells were treated with caspase activating compound doxorubicin (200 ng/ml). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using fluorescence microscope at an interval of 10 min as described in the 'Materials and Methods'. The snapshot of merged channels of ECFP/EYFP/DsRed at 3 h, 6 h, 12 h and 24 h of treatment is shown. The caspase activation is reflected from the FRET loss and subsequent increase in blue channel fluorescence. ( c ) Images representing three distinct cell populations identified after doxorubicin (200 ng/ml) treatment of U251 DEVD Mito-DsRed cell: apoptotic cells with ECFP/EYFP ratio change and retaining red fluorescence, necrotic cells without any ECFP/EYFP fluorescence and retaining red fluorescence and live cells with intact ECFP/EYFP probe without ratio change and retaining red fluorescence. In the image, 'A' represents apoptotic cells, 'N' represents necrotic cells and 'L' represents the live intact cells. ( d ) U251 DEVD Mito Ds cells were either untreated or treated with caspase activating compound for 24 h. The cells were imaged with fluorescence microscope for ratio imaging and DsRed channel. The cells with caspase activation and necrosis were identified on the basis of ratio change and DsRed fluorescence as described. The quantitative necrosis and apoptosis from three different independent experiments are shown in the graph. The results represented are mean±S.D. ( n =3).

    Article Snippet: Expression vector for Mito-DsRed (#6975-1) was purchased from Clontech Laboratories Inc, Mountain View, CA, USA.

    Techniques: Transfection, Expressing, Imaging, Activation Assay, Fluorescence, Microscopy

    ( a ) U251 DEVD Mito Ds cells were treated with CCCP (5 μ M). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using confocal microscopy at an interval of 15 min as described in 'Materials and Methods'. The representative ECFP/EYFP ratio images and merged channels of ECFP/EYFP/DsRed are shown from indicated time points. ( b ) U251 DEVD Mito Ds cells were treated with necrosis-inducing agent H 2 O 2 (2.5 mM). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using confocal microscopy at an interval of 15 min as described in 'Materials and Methods'. The representative ECFP/EYFP ratio images and merged channels of ECFP/EYFP/DsRed are shown from indicated time points.

    Journal: Cell Death Discovery

    Article Title: A quantitative real-time approach for discriminating apoptosis and necrosis

    doi: 10.1038/cddiscovery.2016.101

    Figure Lengend Snippet: ( a ) U251 DEVD Mito Ds cells were treated with CCCP (5 μ M). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using confocal microscopy at an interval of 15 min as described in 'Materials and Methods'. The representative ECFP/EYFP ratio images and merged channels of ECFP/EYFP/DsRed are shown from indicated time points. ( b ) U251 DEVD Mito Ds cells were treated with necrosis-inducing agent H 2 O 2 (2.5 mM). The real-time imaging for caspase activation and Mito-DsRed fluorescence was carried out using confocal microscopy at an interval of 15 min as described in 'Materials and Methods'. The representative ECFP/EYFP ratio images and merged channels of ECFP/EYFP/DsRed are shown from indicated time points.

    Article Snippet: Expression vector for Mito-DsRed (#6975-1) was purchased from Clontech Laboratories Inc, Mountain View, CA, USA.

    Techniques: Imaging, Activation Assay, Fluorescence, Confocal Microscopy

    The schema summarizes the potentiality of the tool to discriminate the two forms of cell death: apoptosis and necrosis in a spatio-temporal manner. The apoptotic cells appear blue due to FRET loss, at the same time retaining the insoluble Mito-DsRed probe; whereas, in necrotic cells, the FRET probe is lost due to membrane permeabilization and are characterized by the presence of insoluble Mito-DsRed probe alone. The healthy cells retain both the intact FRET probe and the Mito-DsRed probe

    Journal: Cell Death Discovery

    Article Title: A quantitative real-time approach for discriminating apoptosis and necrosis

    doi: 10.1038/cddiscovery.2016.101

    Figure Lengend Snippet: The schema summarizes the potentiality of the tool to discriminate the two forms of cell death: apoptosis and necrosis in a spatio-temporal manner. The apoptotic cells appear blue due to FRET loss, at the same time retaining the insoluble Mito-DsRed probe; whereas, in necrotic cells, the FRET probe is lost due to membrane permeabilization and are characterized by the presence of insoluble Mito-DsRed probe alone. The healthy cells retain both the intact FRET probe and the Mito-DsRed probe

    Article Snippet: Expression vector for Mito-DsRed (#6975-1) was purchased from Clontech Laboratories Inc, Mountain View, CA, USA.

    Techniques: