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p5e unc503  (Addgene inc)

 
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    Structured Review

    Addgene inc p5e unc503
    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( <t>unc503:gal4VP16</t> ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).
    P5e Unc503, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Directionality of developing skeletal muscles is set by mechanical forces"

    Article Title: Directionality of developing skeletal muscles is set by mechanical forces

    Journal: Nature Communications

    doi: 10.1038/s41467-023-38647-7

    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( unc503:gal4VP16 ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).
    Figure Legend Snippet: a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( unc503:gal4VP16 ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).

    Techniques Used: Imaging



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    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( <t>unc503:gal4VP16</t> ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).
    P5e Unc503, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( <t>unc503:gal4VP16</t> ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).
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    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( <t>unc503:gal4VP16</t> ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).
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    Image Search Results


    a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( unc503:gal4VP16 ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).

    Journal: Nature Communications

    Article Title: Directionality of developing skeletal muscles is set by mechanical forces

    doi: 10.1038/s41467-023-38647-7

    Figure Lengend Snippet: a Embryos from the outcross between Tg( actb2:loxp:sstop:loxp-DsRed ) and Tg( fli:GFP, tbx1:Cre ) were subjected to live imaging. See also Supplementary Movie . b Differentiating muscle ( tbx1:CreERT2 ) and condensing cartilage ( fli:GFP ) cells were tracked as in ( a ) in control and perturbed embryos. The plots show the extent of polarization in cell shape in ( c ) and the orientation of muscle cells ( d ) relative to pq cartilage. Primary polarized myoblast corresponds to polarized myoblasts adjacent to pq cartilage in the early stage. See also Supplementary Movie . Control 40hpf, n = 28 cells. Control 48hpf, n = 33 cells. Control 56hpf, n = 21 cells. Sox9 -crispant 48hpf, n = 15 cells. Sox9 -crispant 56hpf, n = 18 cells. Runx2b -morphant 48hpf, n = 16 cells. Runx2b -morphant 56hpf, n = 18 cells. Statistical significance in ( c ) was determined by two-way ANOVA ( **** P < 0.0001) e , f The relations between polarity of cells and the distance from pq cartilage in control embryo. Images are 10 μm oblique slices ( e ). Graphs shows scatter plot with best fit power trendline ( f ). 33 cells were measured. The distance was defined as length between the center of pq cartilage and the closest point of cells to it or center of nuclei. See also Supplementary Movie . g The interface between am muscle and pq cartilage was ablated by laser pulse in the embryo from an outcross of Tg( unc503:gal4VP16 ) and Tg( col2:mCherry, UAS:GFP ) around 54hpf. The area of ablation is shown with brackets. The time when the laser-pulse was started is set to 00”00. h The distance from the tip of the muscle cell (light blue arrowhead on ( g )) from the ablated spot was measured and shown. Values are means of 10 embryos ±SD. See also Supplementary Movie . i The Pq pre-cartilage condensation of a 42hpf embryo was laser-ablated, and the polarization of muscle precursor cells were examined with live imaging. The area encompassed by the dashed line was the ablated region. See also Supplementary Movie . j The trajectory of nuclei in differentiating muscle cells in a control embryo. See also Supplementary Movie . k Cell division and the fate of daughter cells (arrowheads) 30 min after cell division were shown in control and cartilage-less embryos. The longitudinal axis of myofibril close to the dividing cells was set as X axis (yellow dash). Images are 3 μm oblique slices. Green dotted line in control shows the division plane. See also Supplementary Movie . Scale bar; 20 μm except in magnified part of ( g ) (5 μm).

    Article Snippet: For the generation of unc503:Gal4VP16, p5E_unc503 (Addgene plasmid # 64020), pME-Gal4VP16 (tol2 kit, #387), and p3E-polyA were recombined to pDESTtol2pACrymCherry (Addgene plasmid # 64023).

    Techniques: Imaging

    Shown are the survival curves for the following promoters and the total number of zebrafish embryos analyzed for each condition: beta actin ( n = 120), mitfa ( n = 45), fli1 ( n = 81), ubi ( n = 160), unc503 ( n = 108), cmv ( n = 20), SA-splice acceptor ( n = 153).

    Journal: eLife

    Article Title: PAX3-FOXO1 transgenic zebrafish models identify HES3 as a mediator of rhabdomyosarcoma tumorigenesis

    doi: 10.7554/eLife.33800

    Figure Lengend Snippet: Shown are the survival curves for the following promoters and the total number of zebrafish embryos analyzed for each condition: beta actin ( n = 120), mitfa ( n = 45), fli1 ( n = 81), ubi ( n = 160), unc503 ( n = 108), cmv ( n = 20), SA-splice acceptor ( n = 153).

    Article Snippet: The ubi promoter was a kind gift from Len Zon (Addgene #27320) , the unc503 promoter from Peter Currie (Addgene #64020) , the fli1 promoter and 3’ entry 2A-mCherry from Nathan Lawson (Addgene #31160 and #26031) ( ; ), and the mitfa promoter from James Lister (Addgene #81234).

    Techniques:

    Journal: eLife

    Article Title: PAX3-FOXO1 transgenic zebrafish models identify HES3 as a mediator of rhabdomyosarcoma tumorigenesis

    doi: 10.7554/eLife.33800

    Figure Lengend Snippet:

    Article Snippet: The ubi promoter was a kind gift from Len Zon (Addgene #27320) , the unc503 promoter from Peter Currie (Addgene #64020) , the fli1 promoter and 3’ entry 2A-mCherry from Nathan Lawson (Addgene #31160 and #26031) ( ; ), and the mitfa promoter from James Lister (Addgene #81234).

    Techniques: Mutagenesis, Injection, Transfection, Construct, Western Blot, Staining, Recombinant, Plasmid Preparation, Generated, Clone Assay, Sequencing, In Situ, Stripping Membranes, Software