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    Lonza shear stress experiment huvecs
    Marcksl1 expression level controls endothelial cell size and shape. a , Live imaging of an ISV composed of EC with ectopic expression of Marcksl1b from 48 hpf reveals changes in cell shape and fluctuation in vessel diameter. Dashed line, region of vessel measured. b - d , Overexpression of Marcksl1 increases EC cell size while depletion of marcksl1b or marcksl1a and marcksl1b decreases EC size in vivo . Methodology to perform in vivo single cell shape analysis ( b ). Quantification of cell area ( c ) and cell aspect ratio ( d ) of lynEGFP-or Marcksl1b-EGFP-expressing ECs surrounded by wildtype (WT) cells, marcksl1 rk23 ; marcksl1b rk24 ECs transplanted in a WT embryo and ECs of marcksl1b rk24 or marcksl1 rk23 ; marcksl1b rk24 embryos at 2 dpf. e , Diameter of ISVs composed of single marcksl1 rk23 ; marcksl1b rk24 EC transplanted into WT embryo at 2 dpf ( n =10 vessels, n =5 independent transplantations). f - m , Overexpression and knockdown of Marcksl1 increases and decreases, respectively, HUVEC size. Maximum intensity projection of confocal z-stacks of control (EGFP) and Marcksl1-overexpressing (Marcksl1-EGFP) <t>HUVECs</t> 1 and 2 days <t>post-transfection</t> (dpt, f ). Quantification of cell area ( g ), cell spikiness index ( h ) and cell aspect ratio ( i ; EGFP: n =423/363 cells at 1 dpt/2 dpt; Marcksl1-EGFP: n = 358/298 cells at 1 dpt/2 dpt). Maximum intensity projection of confocal z-stacks of control and MARCKSL1 knockdown HUVECs stained for DAPI and phalloidin at 1 and 2 dpt ( j ). Plasmid expressing U6 promoter-driven MARCKSL1 shRNA was used with CMV promoter-driven EGFP-CAAX expression as an internal marker. Plasmid expressing scrambled shRNA was used as a control. Quantification of cell area ( k ), cell spikiness index ( l ) and cell aspect ratio ( m ) after MARCKLS1 knockdown (shControl, n =223/278 at 1 dpt/2 dpt cell cultures; shMARCKSL1, n =474/342 at 1 dpt/2 dpt). Statistical significance was assessed by ordinary one-way ANOVA with Tukey’s multiple comparisons test (c and d) and Mann-Whitney U test (e, g-i, k-m). * P
    Shear Stress Experiment Huvecs, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Thermo Fisher shear stress experiment huvecs
    Marcksl1 expression level controls endothelial cell size and shape. a , Live imaging of an ISV composed of EC with ectopic expression of Marcksl1b from 48 hpf reveals changes in cell shape and fluctuation in vessel diameter. Dashed line, region of vessel measured. b - d , Overexpression of Marcksl1 increases EC cell size while depletion of marcksl1b or marcksl1a and marcksl1b decreases EC size in vivo . Methodology to perform in vivo single cell shape analysis ( b ). Quantification of cell area ( c ) and cell aspect ratio ( d ) of lynEGFP-or Marcksl1b-EGFP-expressing ECs surrounded by wildtype (WT) cells, marcksl1 rk23 ; marcksl1b rk24 ECs transplanted in a WT embryo and ECs of marcksl1b rk24 or marcksl1 rk23 ; marcksl1b rk24 embryos at 2 dpf. e , Diameter of ISVs composed of single marcksl1 rk23 ; marcksl1b rk24 EC transplanted into WT embryo at 2 dpf ( n =10 vessels, n =5 independent transplantations). f - m , Overexpression and knockdown of Marcksl1 increases and decreases, respectively, HUVEC size. Maximum intensity projection of confocal z-stacks of control (EGFP) and Marcksl1-overexpressing (Marcksl1-EGFP) <t>HUVECs</t> 1 and 2 days <t>post-transfection</t> (dpt, f ). Quantification of cell area ( g ), cell spikiness index ( h ) and cell aspect ratio ( i ; EGFP: n =423/363 cells at 1 dpt/2 dpt; Marcksl1-EGFP: n = 358/298 cells at 1 dpt/2 dpt). Maximum intensity projection of confocal z-stacks of control and MARCKSL1 knockdown HUVECs stained for DAPI and phalloidin at 1 and 2 dpt ( j ). Plasmid expressing U6 promoter-driven MARCKSL1 shRNA was used with CMV promoter-driven EGFP-CAAX expression as an internal marker. Plasmid expressing scrambled shRNA was used as a control. Quantification of cell area ( k ), cell spikiness index ( l ) and cell aspect ratio ( m ) after MARCKLS1 knockdown (shControl, n =223/278 at 1 dpt/2 dpt cell cultures; shMARCKSL1, n =474/342 at 1 dpt/2 dpt). Statistical significance was assessed by ordinary one-way ANOVA with Tukey’s multiple comparisons test (c and d) and Mann-Whitney U test (e, g-i, k-m). * P
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    Cell Applications Inc shear stress experiments huvecs
    Marcksl1 expression level controls endothelial cell size and shape. a , Live imaging of an ISV composed of EC with ectopic expression of Marcksl1b from 48 hpf reveals changes in cell shape and fluctuation in vessel diameter. Dashed line, region of vessel measured. b - d , Overexpression of Marcksl1 increases EC cell size while depletion of marcksl1b or marcksl1a and marcksl1b decreases EC size in vivo . Methodology to perform in vivo single cell shape analysis ( b ). Quantification of cell area ( c ) and cell aspect ratio ( d ) of lynEGFP-or Marcksl1b-EGFP-expressing ECs surrounded by wildtype (WT) cells, marcksl1 rk23 ; marcksl1b rk24 ECs transplanted in a WT embryo and ECs of marcksl1b rk24 or marcksl1 rk23 ; marcksl1b rk24 embryos at 2 dpf. e , Diameter of ISVs composed of single marcksl1 rk23 ; marcksl1b rk24 EC transplanted into WT embryo at 2 dpf ( n =10 vessels, n =5 independent transplantations). f - m , Overexpression and knockdown of Marcksl1 increases and decreases, respectively, HUVEC size. Maximum intensity projection of confocal z-stacks of control (EGFP) and Marcksl1-overexpressing (Marcksl1-EGFP) <t>HUVECs</t> 1 and 2 days <t>post-transfection</t> (dpt, f ). Quantification of cell area ( g ), cell spikiness index ( h ) and cell aspect ratio ( i ; EGFP: n =423/363 cells at 1 dpt/2 dpt; Marcksl1-EGFP: n = 358/298 cells at 1 dpt/2 dpt). Maximum intensity projection of confocal z-stacks of control and MARCKSL1 knockdown HUVECs stained for DAPI and phalloidin at 1 and 2 dpt ( j ). Plasmid expressing U6 promoter-driven MARCKSL1 shRNA was used with CMV promoter-driven EGFP-CAAX expression as an internal marker. Plasmid expressing scrambled shRNA was used as a control. Quantification of cell area ( k ), cell spikiness index ( l ) and cell aspect ratio ( m ) after MARCKLS1 knockdown (shControl, n =223/278 at 1 dpt/2 dpt cell cultures; shMARCKSL1, n =474/342 at 1 dpt/2 dpt). Statistical significance was assessed by ordinary one-way ANOVA with Tukey’s multiple comparisons test (c and d) and Mann-Whitney U test (e, g-i, k-m). * P
    Shear Stress Experiments Huvecs, supplied by Cell Applications Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Marcksl1 expression level controls endothelial cell size and shape. a , Live imaging of an ISV composed of EC with ectopic expression of Marcksl1b from 48 hpf reveals changes in cell shape and fluctuation in vessel diameter. Dashed line, region of vessel measured. b - d , Overexpression of Marcksl1 increases EC cell size while depletion of marcksl1b or marcksl1a and marcksl1b decreases EC size in vivo . Methodology to perform in vivo single cell shape analysis ( b ). Quantification of cell area ( c ) and cell aspect ratio ( d ) of lynEGFP-or Marcksl1b-EGFP-expressing ECs surrounded by wildtype (WT) cells, marcksl1 rk23 ; marcksl1b rk24 ECs transplanted in a WT embryo and ECs of marcksl1b rk24 or marcksl1 rk23 ; marcksl1b rk24 embryos at 2 dpf. e , Diameter of ISVs composed of single marcksl1 rk23 ; marcksl1b rk24 EC transplanted into WT embryo at 2 dpf ( n =10 vessels, n =5 independent transplantations). f - m , Overexpression and knockdown of Marcksl1 increases and decreases, respectively, HUVEC size. Maximum intensity projection of confocal z-stacks of control (EGFP) and Marcksl1-overexpressing (Marcksl1-EGFP) HUVECs 1 and 2 days post-transfection (dpt, f ). Quantification of cell area ( g ), cell spikiness index ( h ) and cell aspect ratio ( i ; EGFP: n =423/363 cells at 1 dpt/2 dpt; Marcksl1-EGFP: n = 358/298 cells at 1 dpt/2 dpt). Maximum intensity projection of confocal z-stacks of control and MARCKSL1 knockdown HUVECs stained for DAPI and phalloidin at 1 and 2 dpt ( j ). Plasmid expressing U6 promoter-driven MARCKSL1 shRNA was used with CMV promoter-driven EGFP-CAAX expression as an internal marker. Plasmid expressing scrambled shRNA was used as a control. Quantification of cell area ( k ), cell spikiness index ( l ) and cell aspect ratio ( m ) after MARCKLS1 knockdown (shControl, n =223/278 at 1 dpt/2 dpt cell cultures; shMARCKSL1, n =474/342 at 1 dpt/2 dpt). Statistical significance was assessed by ordinary one-way ANOVA with Tukey’s multiple comparisons test (c and d) and Mann-Whitney U test (e, g-i, k-m). * P

    Journal: bioRxiv

    Article Title: Marcksl1 modulates endothelial cell mechanoresponse to haemodynamic forces to control blood vessel shape and size

    doi: 10.1101/2020.08.06.237305

    Figure Lengend Snippet: Marcksl1 expression level controls endothelial cell size and shape. a , Live imaging of an ISV composed of EC with ectopic expression of Marcksl1b from 48 hpf reveals changes in cell shape and fluctuation in vessel diameter. Dashed line, region of vessel measured. b - d , Overexpression of Marcksl1 increases EC cell size while depletion of marcksl1b or marcksl1a and marcksl1b decreases EC size in vivo . Methodology to perform in vivo single cell shape analysis ( b ). Quantification of cell area ( c ) and cell aspect ratio ( d ) of lynEGFP-or Marcksl1b-EGFP-expressing ECs surrounded by wildtype (WT) cells, marcksl1 rk23 ; marcksl1b rk24 ECs transplanted in a WT embryo and ECs of marcksl1b rk24 or marcksl1 rk23 ; marcksl1b rk24 embryos at 2 dpf. e , Diameter of ISVs composed of single marcksl1 rk23 ; marcksl1b rk24 EC transplanted into WT embryo at 2 dpf ( n =10 vessels, n =5 independent transplantations). f - m , Overexpression and knockdown of Marcksl1 increases and decreases, respectively, HUVEC size. Maximum intensity projection of confocal z-stacks of control (EGFP) and Marcksl1-overexpressing (Marcksl1-EGFP) HUVECs 1 and 2 days post-transfection (dpt, f ). Quantification of cell area ( g ), cell spikiness index ( h ) and cell aspect ratio ( i ; EGFP: n =423/363 cells at 1 dpt/2 dpt; Marcksl1-EGFP: n = 358/298 cells at 1 dpt/2 dpt). Maximum intensity projection of confocal z-stacks of control and MARCKSL1 knockdown HUVECs stained for DAPI and phalloidin at 1 and 2 dpt ( j ). Plasmid expressing U6 promoter-driven MARCKSL1 shRNA was used with CMV promoter-driven EGFP-CAAX expression as an internal marker. Plasmid expressing scrambled shRNA was used as a control. Quantification of cell area ( k ), cell spikiness index ( l ) and cell aspect ratio ( m ) after MARCKLS1 knockdown (shControl, n =223/278 at 1 dpt/2 dpt cell cultures; shMARCKSL1, n =474/342 at 1 dpt/2 dpt). Statistical significance was assessed by ordinary one-way ANOVA with Tukey’s multiple comparisons test (c and d) and Mann-Whitney U test (e, g-i, k-m). * P

    Article Snippet: Transfection, siRNA-mediated protein knockdown and shear stress experiment HUVECs (Lonza, C-2519A) were cultured in EGM medium (Lonza) and used until passage 4.

    Techniques: Expressing, Imaging, Over Expression, In Vivo, Transfection, Staining, Plasmid Preparation, shRNA, Marker, MANN-WHITNEY

    Marcksl1 regulates actin organisation in the endothelial cell cortex. a - c , EC cortex contains a dynamic meshwork of actomyosin. Maximum intensity projection of Airyscan images of actin in EC from an ISV of a 2 dpf Tg(fli1ep:GAL4FF) ubs3 ; Tg(UAS:EGFP-UCHD) ubs18 embryo ( a ) and the apical cortex of HUVEC stained with Phalloidin ( b ). Time-lapse imaging of actin and non-muscle myosin II in EC of 2 dpf Tg(fli1:Lifeact-mCherry) ncv7 ; Tg(fli1ep:EGFP-myl9b) rk25 embryo ( c ). Lifeact and Myl9b intensity in boxed region was quantified. d , Maximum intensity projection of 2 z-slices from the apical cortex of HUVEC expressing Marcksl1-EGFP and stained with Phalloidin reveals co-localisation of Marcksl1 and actin bundles (arrowheads). e - i , Analysis of actin density and bundle width after overexpression of Marcksl1, Marcksl1-AAA and Marcksl1-DDD ( e - g ) and knockdown of MARCKSL11 ( h and i ) in HUVECs. Single slice Fast Airyscan images of the apical cortex of HUVEC stained with Phalloidin reveal decreased actin density surrounding actin bundles (arrow) in Marcksl1-EGFP-transfected cells 1 day post transfection ( e ). f - i , Mean values are indicated. 3 independent experiments were performed (EGFP, 51 ROIs from 28 cells; Marcksl1-EGFP, 86 ROIs from 29 cells; Marcksl1-AAA-EGFP, 76 ROIs from 28 cells; Marcksl1-DDD-EGFP, 77 ROIs from 32 cells; shControl, 66 ROIs from 24 cells; shMARCKSL1, 74 ROIs from 29 cells). Data was analysed by Mann-Whitney test. **** P

    Journal: bioRxiv

    Article Title: Marcksl1 modulates endothelial cell mechanoresponse to haemodynamic forces to control blood vessel shape and size

    doi: 10.1101/2020.08.06.237305

    Figure Lengend Snippet: Marcksl1 regulates actin organisation in the endothelial cell cortex. a - c , EC cortex contains a dynamic meshwork of actomyosin. Maximum intensity projection of Airyscan images of actin in EC from an ISV of a 2 dpf Tg(fli1ep:GAL4FF) ubs3 ; Tg(UAS:EGFP-UCHD) ubs18 embryo ( a ) and the apical cortex of HUVEC stained with Phalloidin ( b ). Time-lapse imaging of actin and non-muscle myosin II in EC of 2 dpf Tg(fli1:Lifeact-mCherry) ncv7 ; Tg(fli1ep:EGFP-myl9b) rk25 embryo ( c ). Lifeact and Myl9b intensity in boxed region was quantified. d , Maximum intensity projection of 2 z-slices from the apical cortex of HUVEC expressing Marcksl1-EGFP and stained with Phalloidin reveals co-localisation of Marcksl1 and actin bundles (arrowheads). e - i , Analysis of actin density and bundle width after overexpression of Marcksl1, Marcksl1-AAA and Marcksl1-DDD ( e - g ) and knockdown of MARCKSL11 ( h and i ) in HUVECs. Single slice Fast Airyscan images of the apical cortex of HUVEC stained with Phalloidin reveal decreased actin density surrounding actin bundles (arrow) in Marcksl1-EGFP-transfected cells 1 day post transfection ( e ). f - i , Mean values are indicated. 3 independent experiments were performed (EGFP, 51 ROIs from 28 cells; Marcksl1-EGFP, 86 ROIs from 29 cells; Marcksl1-AAA-EGFP, 76 ROIs from 28 cells; Marcksl1-DDD-EGFP, 77 ROIs from 32 cells; shControl, 66 ROIs from 24 cells; shMARCKSL1, 74 ROIs from 29 cells). Data was analysed by Mann-Whitney test. **** P

    Article Snippet: Transfection, siRNA-mediated protein knockdown and shear stress experiment HUVECs (Lonza, C-2519A) were cultured in EGM medium (Lonza) and used until passage 4.

    Techniques: Staining, Imaging, Expressing, Over Expression, Transfection, MANN-WHITNEY