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  • 93
    Addgene inc aktar2
    All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of <t>AktAR2</t> FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.
    Aktar2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cyagen Biosciences cmv promoter
    All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of <t>AktAR2</t> FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.
    Cmv Promoter, supplied by Cyagen Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cmv promoter/product/Cyagen Biosciences
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    93
    Santa Cruz Biotechnology control sirna
    All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of <t>AktAR2</t> FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.
    Control Sirna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher lipofectamine 2000
    All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of <t>AktAR2</t> FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.
    Lipofectamine 2000, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of AktAR2 FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.

    Journal: Nature Communications

    Article Title: Optogenetic regulation of endogenous proteins

    doi: 10.1038/s41467-020-14460-4

    Figure Lengend Snippet: All-optical control of endogenous RAS in steady state. a RAS regulation by light in cells expressing EKAR2G2, in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0 and white = 2.05. b Normalized whole-cell average FRET/donor ratio in HeLa cells expressing the optogenetically enhanced iB(RAS) and EKAR2G2 FRET biosensor. n = 3, error bars represent SEM. Source data are provided as a Source Data file. c Illumination scheme used for the experiments shown in Fig. 9. d RAS-Akt signaling regulation in cells expressing optogenetically controlled iB(RAS). Representative time-lapse panels of AktAR2 FRET/donor ratio, imaged in HeLa cells in serum. Epifluorescence microscopy; scale bar, 20 µm. Pseudocolor scale: black = 1.0; white = 2.88. e Normalized whole-cell average FRET/donor ratio as a function of time, in HeLa cells expressing the AktAR2 FRET biosensor. Error bars represent SEM, n = 3 independent experiments. Source data are provided as a Source Data file. b , e Black: control cells without light-activation. Green: cells with 740 nm illumination starting at t = 300 s time point. Magenta: cells were irradiated with 740 nm light for 2400 s prior to imaging. f Schematic representation of all-optical control of RAS signaling using iB(RAS). Signaling nodes are shown in gray, optogenetic tools and biosensors are shown in light gray. Fast negative feedback loops acting via phosphorylation are shown as red dotted lines. Slow feedback loops acting via transcription inhibition are shown in gray.

    Article Snippet: For EKAR2G2 (Addgene #40178) and AKTAR2 (Addgene #64932) biosensor experiments, HeLa cells were transiently transfected together with pQP-AR10 (Supplementary Table ) construct at DNA ratio of 1: 1.

    Techniques: Expressing, Epifluorescence Microscopy, Activation Assay, Irradiation, Imaging, Inhibition