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  • 89
    JEOL 100 cx electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    100 Cx Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 89/100, based on 458 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    JEOL jem 100 cx electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Jem 100 Cx Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 88/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    JEOL cx 100 scanning electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Cx 100 Scanning Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    JEOL jsm 100 cx transmission electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Jsm 100 Cx Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    JEOL jsm 100 cx 11 electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Jsm 100 Cx 11 Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    JEOL model jem 100 cx transmission electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Model Jem 100 Cx Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    JEOL 100 cx ii 100kw transmission electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    100 Cx Ii 100kw Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 10 article reviews
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    80
    JEOL model 100 cx transmission electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Model 100 Cx Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 80/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    JEOL japanese electron optical laboratories 100 cx transmission electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    Japanese Electron Optical Laboratories 100 Cx Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; 100 mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.

    Journal: Genes & Development

    Article Title: members only encodes a Drosophila nucleoporin required for Rel protein import and immune response activation

    doi:

    Figure Lengend Snippet: The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; 100 mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.

    Article Snippet: Wild-type and mbo mutant larvae were prepared for EM as described ( ) and examined with a Jeol 100 CX electron microscope.

    Techniques: Expressing, Staining, In Situ Hybridization, Mutagenesis, Marker

    mbo is not required for mRNA export. ( A–C ) In situ hybridization to lacZ RNA in wild-type and mbo larvae carrying the hs–GAL4 and UAS–lacZNLS transgenes. The lacZ RNA is detected in the proventriculus of wild-type ( B ) and mbo mutant ( C ) larvae after heat shock and does not accumulate in the nucleus (arrowheads). The dark spot inside each nucleus is likely to correlate with the site of transcription. lacZ expression is reduced in some of the cells of mbo mutants (arrows). Bar, 10 μm. ( D–F ) Heat shock-induced expression of Hdc protein in wild-type and mbo mutants. Fat bodies from untreated wild-type ( D ) and heat-shocked wild-type ( E ) and mbo ( F ) larvae carrying the hs–hdc transgene were stained with an antibody against the Hdc protein. Bar, 50 μm. ( G ) Electron micrograph of a section through the lymph gland of an mbo larva. In this tangential section, several NPCs (arrow) can be identified in the space between the cytoplasm (Cyt) and the nucleus (Nuc). Their distribution and morphology are indistinguishable from wild type at this level. Bar, 100 nm.

    Journal: Genes & Development

    Article Title: members only encodes a Drosophila nucleoporin required for Rel protein import and immune response activation

    doi:

    Figure Lengend Snippet: mbo is not required for mRNA export. ( A–C ) In situ hybridization to lacZ RNA in wild-type and mbo larvae carrying the hs–GAL4 and UAS–lacZNLS transgenes. The lacZ RNA is detected in the proventriculus of wild-type ( B ) and mbo mutant ( C ) larvae after heat shock and does not accumulate in the nucleus (arrowheads). The dark spot inside each nucleus is likely to correlate with the site of transcription. lacZ expression is reduced in some of the cells of mbo mutants (arrows). Bar, 10 μm. ( D–F ) Heat shock-induced expression of Hdc protein in wild-type and mbo mutants. Fat bodies from untreated wild-type ( D ) and heat-shocked wild-type ( E ) and mbo ( F ) larvae carrying the hs–hdc transgene were stained with an antibody against the Hdc protein. Bar, 50 μm. ( G ) Electron micrograph of a section through the lymph gland of an mbo larva. In this tangential section, several NPCs (arrow) can be identified in the space between the cytoplasm (Cyt) and the nucleus (Nuc). Their distribution and morphology are indistinguishable from wild type at this level. Bar, 100 nm.

    Article Snippet: Wild-type and mbo mutant larvae were prepared for EM as described ( ) and examined with a Jeol 100 CX electron microscope.

    Techniques: In Situ Hybridization, Mutagenesis, Expressing, Staining

    Nucleotide effects on RME-1 and AMPH-1 mediated liposome tubulation. (a) Coomassie stained gels of supernatant and pellet fractions from liposome co-sedimentation assays are shown. Binding reactions were performed in the absence or presence of 0.33 mg/ml, 0.4 µm (average diameter) 100% Phosphatidylserine (PtdSer), or 100% Phosphatidylcholine (PtdChl) liposomes. Liposomes were incubated with 1mM ADP and 1 µM of full length AMPH-1 or RME-1 proteins, or equimolar quantities of both proteins, as indicated. Note that RME-1 can bind to PtdSer liposomes in the presence of ADP. Tubulation experiments were performed with 2.5µM of each protein and 1mM ADP incubated with 0.05mg/ml 100% PtdSer liposomes. (b-b’) AMPH-1 incubated with PtdSer liposomes in the presence of ADP (compare with ATP-γ-S, Fig. 5 c-c’ ). (c-c’) RME-1 incubated with PtdSer liposomes in the presence of ADP (compare with ATP-γ-S, Fig. 5 d-d’ ). RME-1(ADP) lacks tubulation capacity and most liposomes remain spherical (open arrow) in the presence of RME-1(ADP). Striations are often visible and rare short protrusions (closed arrows) are present on occasional liposomes. (d-d’) RME-1 in its ADP bound state can affect the tubulation ability of AMPH-1 as observed in an experiment containing equimolar concentrations of both proteins in the presence of ADP. (e) Quantification reveals an approximately 7 fold decrease in number of tubules in conditions where RME-1 is present with AMPH-1, as compared to tubulation produced by AMPH-1 alone. n=10 fields for each experimental condition (imaged at 3,000× magnification), the mean value for number of observed tubules was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test (p value=1.48×10 −15 ).

    Journal: Nature cell biology

    Article Title: AMPH-1/Amphiphysin/Bin1 functions with RME-1/Ehd in endocytic recycling

    doi: 10.1038/ncb1986

    Figure Lengend Snippet: Nucleotide effects on RME-1 and AMPH-1 mediated liposome tubulation. (a) Coomassie stained gels of supernatant and pellet fractions from liposome co-sedimentation assays are shown. Binding reactions were performed in the absence or presence of 0.33 mg/ml, 0.4 µm (average diameter) 100% Phosphatidylserine (PtdSer), or 100% Phosphatidylcholine (PtdChl) liposomes. Liposomes were incubated with 1mM ADP and 1 µM of full length AMPH-1 or RME-1 proteins, or equimolar quantities of both proteins, as indicated. Note that RME-1 can bind to PtdSer liposomes in the presence of ADP. Tubulation experiments were performed with 2.5µM of each protein and 1mM ADP incubated with 0.05mg/ml 100% PtdSer liposomes. (b-b’) AMPH-1 incubated with PtdSer liposomes in the presence of ADP (compare with ATP-γ-S, Fig. 5 c-c’ ). (c-c’) RME-1 incubated with PtdSer liposomes in the presence of ADP (compare with ATP-γ-S, Fig. 5 d-d’ ). RME-1(ADP) lacks tubulation capacity and most liposomes remain spherical (open arrow) in the presence of RME-1(ADP). Striations are often visible and rare short protrusions (closed arrows) are present on occasional liposomes. (d-d’) RME-1 in its ADP bound state can affect the tubulation ability of AMPH-1 as observed in an experiment containing equimolar concentrations of both proteins in the presence of ADP. (e) Quantification reveals an approximately 7 fold decrease in number of tubules in conditions where RME-1 is present with AMPH-1, as compared to tubulation produced by AMPH-1 alone. n=10 fields for each experimental condition (imaged at 3,000× magnification), the mean value for number of observed tubules was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test (p value=1.48×10 −15 ).

    Article Snippet: After negative staining with 1% uranyl acetate, electron microscopy was performed using a JEOL 1200 EX or JEOL 100 CX transmission electron microscope at 80 KV.

    Techniques: Staining, Sedimentation, Binding Assay, Incubation, Produced, One-tailed Test, Significance Assay

    Membrane binding and tubulation by AMPH-1 and RME-1 in vitro . ( a ) Coomassie stained gels of supernatant and pellet fractions from liposome co-sedimentation assays are shown. Binding reactions were performed in the absence or presence of 0.33 mg/ml, 0.4 µm (average diameter) 100% Phosphatidylserine (PtdSer), or 100% Phosphatidylcholine (PtdChl) liposomes. Liposomes were incubated with 1mM ATP-γ-S and 1 µM of full length AMPH-1 or RME-1 proteins, or equimolar quantities of both proteins, as indicated. ( b-e’ ) Electron micrographs of negatively stained PtdSer liposomes, prepared as above, but used at 0.05 mg/ml in the presence of 1mM ATP-γ-S with 2.5 µM proteins, GST ( b-b’ ), AMPH-1 ( c-c’ ), RME-1 ( d-d’ ), or equimolar quantities of AMPH-1 and RME-1 ( e-e’ ). ( f ) Quantification of tubule widths. For each experimental condition, width was measured for every tubule on each of 15 electron micrographs. For tubules demonstrating uneven width, an average measurement was made after taking measures at several representative points along the tubule. For n=15 representative tubules per experimental condition, the mean value for tubule width was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test: RME-1 vs. RME-1+AMPH-1, p value= 0.0074 and for AMPH-1 vs. RME-1+AMPH-1, p value= 1.29 × 10 −13 . ( g ) Quantification of tubule lengths. The length from the edge of the liposome body to end of tubule was measured for every tubule on each of 15 electron micrographs per experimental condition. For n=15 representative tubules for each condition, the mean value for tubule length was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test for tubule length: RME-1 vs. RME-1+AMPH-1, p value=1.2 × 10 −07 and for AMPH-1 vs. RME-1+AMPH-1, p value= 3.2 × 10 −04 . ( h ) Quantification of inter-striation distance. For n=15 representative tubules, the average distance between every successive ring-like striation was counted for each experimental condition. Mean values were plotted and error bars represent ± standard deviation from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test, p value=0.0031.

    Journal: Nature cell biology

    Article Title: AMPH-1/Amphiphysin/Bin1 functions with RME-1/Ehd in endocytic recycling

    doi: 10.1038/ncb1986

    Figure Lengend Snippet: Membrane binding and tubulation by AMPH-1 and RME-1 in vitro . ( a ) Coomassie stained gels of supernatant and pellet fractions from liposome co-sedimentation assays are shown. Binding reactions were performed in the absence or presence of 0.33 mg/ml, 0.4 µm (average diameter) 100% Phosphatidylserine (PtdSer), or 100% Phosphatidylcholine (PtdChl) liposomes. Liposomes were incubated with 1mM ATP-γ-S and 1 µM of full length AMPH-1 or RME-1 proteins, or equimolar quantities of both proteins, as indicated. ( b-e’ ) Electron micrographs of negatively stained PtdSer liposomes, prepared as above, but used at 0.05 mg/ml in the presence of 1mM ATP-γ-S with 2.5 µM proteins, GST ( b-b’ ), AMPH-1 ( c-c’ ), RME-1 ( d-d’ ), or equimolar quantities of AMPH-1 and RME-1 ( e-e’ ). ( f ) Quantification of tubule widths. For each experimental condition, width was measured for every tubule on each of 15 electron micrographs. For tubules demonstrating uneven width, an average measurement was made after taking measures at several representative points along the tubule. For n=15 representative tubules per experimental condition, the mean value for tubule width was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test: RME-1 vs. RME-1+AMPH-1, p value= 0.0074 and for AMPH-1 vs. RME-1+AMPH-1, p value= 1.29 × 10 −13 . ( g ) Quantification of tubule lengths. The length from the edge of the liposome body to end of tubule was measured for every tubule on each of 15 electron micrographs per experimental condition. For n=15 representative tubules for each condition, the mean value for tubule length was plotted and error bars represent ± s.d. from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test for tubule length: RME-1 vs. RME-1+AMPH-1, p value=1.2 × 10 −07 and for AMPH-1 vs. RME-1+AMPH-1, p value= 3.2 × 10 −04 . ( h ) Quantification of inter-striation distance. For n=15 representative tubules, the average distance between every successive ring-like striation was counted for each experimental condition. Mean values were plotted and error bars represent ± standard deviation from the mean. The asterisk indicates a significant difference in the one-tailed Student’s T-test, p value=0.0031.

    Article Snippet: After negative staining with 1% uranyl acetate, electron microscopy was performed using a JEOL 1200 EX or JEOL 100 CX transmission electron microscope at 80 KV.

    Techniques: Binding Assay, In Vitro, Staining, Sedimentation, Incubation, One-tailed Test, Standard Deviation

    Characterization of PEGylated AuNRs (length 26±3 nm, width 5±0.8 nm). Notes: ( A ) TEM image with 100 nm scale bar. ( B ) UV–Vis absorbance spectra showing the SPR peaks of AuNRs. ( C ) Corresponding histograms of the lengths of the 100 AuNRs particles counted. ( D ) Corresponding histograms of the widths of the 100 AuNRs particles counted. ( E ) Zeta potential data of the as-synthesized rods. ( F ) Zeta potential data of PEGylated AuNRs. Abbreviations: AuNRs, gold nanorods; TEM, transmission electron microscope; SPR, surface plasmon resonance; UV–Vis, ulltraviolet–visible.

    Journal: International Journal of Nanomedicine

    Article Title: Treatment of natural mammary gland tumors in canines and felines using gold nanorods-assisted plasmonic photothermal therapy to induce tumor apoptosis

    doi: 10.2147/IJN.S109470

    Figure Lengend Snippet: Characterization of PEGylated AuNRs (length 26±3 nm, width 5±0.8 nm). Notes: ( A ) TEM image with 100 nm scale bar. ( B ) UV–Vis absorbance spectra showing the SPR peaks of AuNRs. ( C ) Corresponding histograms of the lengths of the 100 AuNRs particles counted. ( D ) Corresponding histograms of the widths of the 100 AuNRs particles counted. ( E ) Zeta potential data of the as-synthesized rods. ( F ) Zeta potential data of PEGylated AuNRs. Abbreviations: AuNRs, gold nanorods; TEM, transmission electron microscope; SPR, surface plasmon resonance; UV–Vis, ulltraviolet–visible.

    Article Snippet: Characterization of AuNRs The characterization of AuNRs was carried out using a Cary 500 UV–Vis Spectrometer (Agilent Technologies, Santa Clara, CA, USA) for the spectroscopic measurements, and a JEOL 100 CX transmission electron microscope (TEM) (JEOL Ltd., Tokyo, Japan) was used to image the samples.

    Techniques: Transmission Electron Microscopy, SPR Assay, Synthesized, Transmission Assay, Microscopy

    Knockdown of SR-BI/CLA-1 in Caco-2/TC7 cells impairs PPM-induced ERK1/2 phosphorylation and apoB chase. (A) Caco-2/TC7 Cell populations 63 and 64, expressing lentiviral shRNA 63 and 64 respectively, were analyzed at passage 4 after transfection in the absence of PPM or after 10 min of PPM supply. Cell lysates were analyzed by immunoblot with antibodies against SR-BI/CLA-1 and E-cadherin (E-cadh, used as loading control). The lower panel shows the level of SR-BI/CLA-1 expression normalized to the level of E-cadherin expression set at 100% for control Caco-2/TC7 cells. Results are from two independent sets of experiments. (B) Cell populations 63 and 64 were cultured on semi-permeable filters and incubated in the absence or presence of PPM or IPM in the apical compartment for the indicated times. An early (63E) and a late (63L) passage (corresponding respectively to passage 6 and 28 after transfection) of Cell population 63 were compared to Cell population 64 at passage 28. Cell lysates were analyzed in SR-BI/CLA-1 and phospho-ERK1/2 (P-ERK) immunoblots. Total ERK (ERK) and E-cadherin (E-cadh) were used as loading controls. Lower panel, the ratio of P-ERK expression normalized to total ERK expression in PPM-treated cells versus IPM-treated cells, set at 100% for Cell population 64. Results show the means±SEM of three independent sets of experiments. *P

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: Knockdown of SR-BI/CLA-1 in Caco-2/TC7 cells impairs PPM-induced ERK1/2 phosphorylation and apoB chase. (A) Caco-2/TC7 Cell populations 63 and 64, expressing lentiviral shRNA 63 and 64 respectively, were analyzed at passage 4 after transfection in the absence of PPM or after 10 min of PPM supply. Cell lysates were analyzed by immunoblot with antibodies against SR-BI/CLA-1 and E-cadherin (E-cadh, used as loading control). The lower panel shows the level of SR-BI/CLA-1 expression normalized to the level of E-cadherin expression set at 100% for control Caco-2/TC7 cells. Results are from two independent sets of experiments. (B) Cell populations 63 and 64 were cultured on semi-permeable filters and incubated in the absence or presence of PPM or IPM in the apical compartment for the indicated times. An early (63E) and a late (63L) passage (corresponding respectively to passage 6 and 28 after transfection) of Cell population 63 were compared to Cell population 64 at passage 28. Cell lysates were analyzed in SR-BI/CLA-1 and phospho-ERK1/2 (P-ERK) immunoblots. Total ERK (ERK) and E-cadherin (E-cadh) were used as loading controls. Lower panel, the ratio of P-ERK expression normalized to total ERK expression in PPM-treated cells versus IPM-treated cells, set at 100% for Cell population 64. Results show the means±SEM of three independent sets of experiments. *P

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Expressing, shRNA, Transfection, Cell Culture, Incubation, Western Blot

    Subcellular localization of SR-BI/CLA-1 after the supply of postprandial micelles. (A) Immunoelectron micrograph of SR-BI/CLA-1 in untreated differentiated Caco-2/TC7 cells. MV, microvilli; TW, terminal web (bar, 0.5 µm). Note the significant amount of intracellular trafficking SR-BI/CLA-1 in addition to its main apical localization (arrowheads). (B) Immunolocalization of SR-BI/CLA-1 (green channel) and sucrase isomaltase (SI, red channel) in differentiated Caco-2/TC7 cells before (T0) and after 5, 10 and 15 min of apical PPM supply. Panels represent XY acquisitions at the apical level (bar, 10 µm). Arrowheads show clusters of SR-BI/CLA-1. (C) Immunolocalization of SR-BI/CLA-1 in differentiated Caco-2/TC7 cells in the absence (control) or presence of PPM or IPM for 20 min (bar, 20 µm). Arrowheads show clusters of SR-BI/CLA-1 (D) Immunoelectron micrograph of SR-BI/CLA-1 in Caco-2/TC7 cells supplied with PPM (MV, microvilli). Arrowheads indicate SR-BI/CLA-1 clusters (bar, 100 nm). (E) Cell surface biotinylation assay for apical SR-BI/CLA-1. Caco-2/TC7 cells were cultured in the absence (0) or presence of PPM for the indicated times. Cells were then selectively labeled with non-permeant biotin at the apical (left panel) or basal surface (right panel). Biotinylated fractions were obtained as described in Material and Methods . Total cell lysates (total), apical and basal biotinylated fractions (left and right panel respectively) and non-apical fractions (non-apical) were analyzed in immunoblots of SR-BI/CLA-1, E-cadherin being used as a basolateral membrane marker.

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: Subcellular localization of SR-BI/CLA-1 after the supply of postprandial micelles. (A) Immunoelectron micrograph of SR-BI/CLA-1 in untreated differentiated Caco-2/TC7 cells. MV, microvilli; TW, terminal web (bar, 0.5 µm). Note the significant amount of intracellular trafficking SR-BI/CLA-1 in addition to its main apical localization (arrowheads). (B) Immunolocalization of SR-BI/CLA-1 (green channel) and sucrase isomaltase (SI, red channel) in differentiated Caco-2/TC7 cells before (T0) and after 5, 10 and 15 min of apical PPM supply. Panels represent XY acquisitions at the apical level (bar, 10 µm). Arrowheads show clusters of SR-BI/CLA-1. (C) Immunolocalization of SR-BI/CLA-1 in differentiated Caco-2/TC7 cells in the absence (control) or presence of PPM or IPM for 20 min (bar, 20 µm). Arrowheads show clusters of SR-BI/CLA-1 (D) Immunoelectron micrograph of SR-BI/CLA-1 in Caco-2/TC7 cells supplied with PPM (MV, microvilli). Arrowheads indicate SR-BI/CLA-1 clusters (bar, 100 nm). (E) Cell surface biotinylation assay for apical SR-BI/CLA-1. Caco-2/TC7 cells were cultured in the absence (0) or presence of PPM for the indicated times. Cells were then selectively labeled with non-permeant biotin at the apical (left panel) or basal surface (right panel). Biotinylated fractions were obtained as described in Material and Methods . Total cell lysates (total), apical and basal biotinylated fractions (left and right panel respectively) and non-apical fractions (non-apical) were analyzed in immunoblots of SR-BI/CLA-1, E-cadherin being used as a basolateral membrane marker.

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Cell Surface Biotinylation Assay, Cell Culture, Labeling, Western Blot, Marker

    PPM supply induces movement of SR-BI/CLA-1 towards raft microdomains. (A) Caco-2/TC7 cells were harvested in the presence of Triton X-100 and the lysate fractionated on a 5–40% sucrose gradient. Eleven fractions were collected for immunoblots of SR-BI/CLA-1, EEA1 (early endosome antigen 1) and flottilin-1 (raft marker). (B) Caco-2/TC7 cells were cultured in the absence (control) or presence of PPM or IPM for 10 min and then harvested in the presence of Triton X-100. Cell lysates were applied to a 5–40% sucrose gradient and eleven fractions collected. Fractions 3 to 8 were analyzed by immunoblotting with antibodies against SR-BI/CLA-1 (left panel) and flottilin-1 (right panel). (C) Immunolocalization of SR-BI/CLA-1 and alkaline phosphatase (PLAP, used as raft marker) in the brush border of Caco-2/TC7 cells supplied with PPM. SR-BI/CLA-1 is labelled with anti-rabbit immunoglobulin-gold complexes (18-nm particles) and PLAP with anti-sheep immunoglobulin-gold complexes (12-nm particles). MV, microvilli; bar, 100 nm.

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: PPM supply induces movement of SR-BI/CLA-1 towards raft microdomains. (A) Caco-2/TC7 cells were harvested in the presence of Triton X-100 and the lysate fractionated on a 5–40% sucrose gradient. Eleven fractions were collected for immunoblots of SR-BI/CLA-1, EEA1 (early endosome antigen 1) and flottilin-1 (raft marker). (B) Caco-2/TC7 cells were cultured in the absence (control) or presence of PPM or IPM for 10 min and then harvested in the presence of Triton X-100. Cell lysates were applied to a 5–40% sucrose gradient and eleven fractions collected. Fractions 3 to 8 were analyzed by immunoblotting with antibodies against SR-BI/CLA-1 (left panel) and flottilin-1 (right panel). (C) Immunolocalization of SR-BI/CLA-1 and alkaline phosphatase (PLAP, used as raft marker) in the brush border of Caco-2/TC7 cells supplied with PPM. SR-BI/CLA-1 is labelled with anti-rabbit immunoglobulin-gold complexes (18-nm particles) and PLAP with anti-sheep immunoglobulin-gold complexes (12-nm particles). MV, microvilli; bar, 100 nm.

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Western Blot, Marker, Cell Culture

    Two-photon microscopy imaging of FM 1–43 to determine synaptic vesicle endocytosis in crayfish and mice. ( A ) ( a ) The fluorescence image of mice motor nerve terminal-loaded FM 1–43 with tetanic stimulation in the presence of 100 μM T-588. Dye loading was repeated after washing out of T-588. Images were captured every 1-μm step in the Z direction and accumulated. The brightest images in the terminal are shown expanded in the lower column. ( b ) After full loading of FM 1–43, the preparation was treated with 100 μM T-588 for 30 min and then destained by tetanic stimulation (20 Hz, 10 min) without dye in the bath in the presence of T-588. The same set of experiments was repeated without T-588. ( B ) Fluorescence image of crayfish motor nerve terminal-loaded FM 1–43 with tetanic stimulation. Dye loading was repeated with 100 μM T-588 and washing out of T-588 after taking under control condition. Scale bar in each image represents 2 μm.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Reduced facilitation and vesicular uptake in crustacean and mammalian neuromuscular junction by T-588, a neuroprotective compound

    doi:

    Figure Lengend Snippet: Two-photon microscopy imaging of FM 1–43 to determine synaptic vesicle endocytosis in crayfish and mice. ( A ) ( a ) The fluorescence image of mice motor nerve terminal-loaded FM 1–43 with tetanic stimulation in the presence of 100 μM T-588. Dye loading was repeated after washing out of T-588. Images were captured every 1-μm step in the Z direction and accumulated. The brightest images in the terminal are shown expanded in the lower column. ( b ) After full loading of FM 1–43, the preparation was treated with 100 μM T-588 for 30 min and then destained by tetanic stimulation (20 Hz, 10 min) without dye in the bath in the presence of T-588. The same set of experiments was repeated without T-588. ( B ) Fluorescence image of crayfish motor nerve terminal-loaded FM 1–43 with tetanic stimulation. Dye loading was repeated with 100 μM T-588 and washing out of T-588 after taking under control condition. Scale bar in each image represents 2 μm.

    Article Snippet: Thin sections from the chosen areas were collected on single slot, Pyoloform, and carbon-coated grids, contrasted with uranyl acetate and lead citrate, and photographed with a Phillips 208 or a JEOL 100 CX electron microscopes.

    Techniques: Microscopy, Imaging, Mouse Assay, Fluorescence