bioimagesUnknownM. Rizwan SiddiquiThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201012129JCB

Z-stack images of Cav-1–/– endothelial monolayers pre-treated with L-NNA and stained for VE-cadherin (green), b-catenin (magenta), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope equipped with c-Apochromat 63x/1.2 water correction objective and the following filter sets: Alexa-488: BP 505/530; Alexa-532: BP 560/615 and Cy5: LP 650.<br />

Z-stack images of Wt endothelial monolayers stained for VE-cadherin (green), beta-catenin (magenta), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope equipped with c-Apochromat 63x/1.2 water correction objective and the following filter sets: Alexa-488: BP 505/530; Alexa-532: BP 560/615 and Cy5: LP 650.

Immunofluorescent staining of HPAECs for eNOS (green) and b-cat (red); DAPI (blue); 3 min after stimulation with 50 nM a-thrombin

VE-cadherin distribution in Wt endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).<br /><br />

Organization of actin cytoskeleton in Cav-1–/– endothelial monolayers: 12-bit projected image.

Confocal microscopy images of control and Ga13 siRNA lentivirus-infected confluent HMVEC-L in response to 300M H2O2, 2g/ml LPS, or 20ng/ml TNFa.

Confocol microscopy images of VE-cadherin subcellular location in mScr or mG13BP pre-treated HMVEC-L in the presence or absence of chloroquine for 3 hr. Green: VE-cadherin. Arrows pointed to internalized VE-cadherin.

Beta-catenin distribution in Cav-1–/– endothelial monolayers pretreated with RhoA inhibitor: 12-bit projected image.

Confocal microscopy images of confluent HMVEC-L pre-treated with PBS, 150uM mScr, or 150uM mG13BP in response to PBS, 300uM H2O2, 2ug/ml LPS, or 20ng/ml TNFa.

Immunofluorescent staining of HPAECs for eNOS (green) and b-cat (red); DAPI (blue); 15 min after stimulation with 50 nM a-thrombin

Organization of actin cytoskeleton in Cav-1–/– endothelial monolayers pre-treated with Rho inhibitor C3 transferase: 12-bit projected image.

Organization of actin cytoskeleton in eNOS–/–/Cav-1–/– double KO endothelial monolayers: 12-bit projected image. <br /><br />

VE-cadherin distribution in Cav-1–/– endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Organization of actin cytoskeleton in Cav-1–/– endothelial monolayers: 16-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).<br />

Z-stack images of Cav-1–/– endothelial monolayers stained for VE-cadherin (green), beta-catenin (magenta), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope equipped with c-Apochromat 63x/1.2 water correction objective and the following filter sets: Alexa-488: BP 505/530; Alexa-532: BP 560/615 and Cy5: LP 650.<br />

Beta-catenin distribution in Cav-1–/– endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).<br />

Immunofluorescent staining of HPAECs for eNOS (green) and b-cat (red); DAPI (blue); untreated cells.

VE-cadherin distribution in Cav-1–/– endothelial monolayers pretreated with L-NNA: 16-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Z-stack images of Cav-1–/– endothelial monolayers pre-treated with AP-CSD peptide and stained for beta-catenin (green), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope.

Organization of actin cytoskeleton in Cav-1–/– endothelial monolayers pretreated with L-NNA: 16-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).<br />

Live cell images of a FRET-based RhoA biosensor in Wt endothelial monolayers. Z-stack images were obtained using a confocal microscope (Zeiss LSM 510 META) equipped with 63X, 1.2 NA water immersion correction objective and an Ar ion laser. For emission ratio imaging, we acquired 12-bit CFP (blue; l=458 nm; BP500/20), FRET (red; l=458 nm; LP530) and YFP (yellow; l=514 nm; LP530) images. <br />

Beta-catenin distribution in eNOS–/–/Cav-1–/– double KO endothelial monolayers: 12-bit projected image. <br />

Organization of actin cytoskeleton in Wt endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Beta-catenin distribution in Wt endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Beta-catenin distribution in Cav-1–/– endothelial monolayers: 12-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Live cell images of a FRET-based RhoA biosensor in Cav-1-/- endothelial monolayers. Z-stack images were obtained using a confocal microscope (Zeiss LSM 510 META) equipped with 63X, 1.2 NA water immersion correction objective and an Ar ion lasers. For emission ratio imaging, we acquired 12-bit CFP (blue; l=458 nm; BP500/20), FRET (red; l=458 nm; LP530) and YFP (yellow; l=514 nm; LP530) images. <br />

Confocal microscopy images of VE-cadherin expression in mouse PMECs isolated from WT and Gna13flox/flox mice infected with Cre recombinase adenovirus.

Z-stack images of eNOS–/–/Cav-1–/– double KO endothelial monolayers stained for beta-catenin (green), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope<br />

Organization of actin cytoskeleton in Cav-1–/– endothelial monolayers pretreated with AP-CSD peptide: 12-bit projected image.

Beta-catenin distribution in Cav-1–/– endothelial monolayers pretreated with L-NNA: 16-bit projected image. The maximum pixel intensity was collected from each image of Z-stack and projected to the single image using MetaMorph software (Molecular Devices).

Z-stack images of Cav-1–/– endothelial monolayers stained for VE-cadherin (green), beta-catenin (magenta), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope equipped with c-Apochromat 63x/1.2 water correction objective.

Z-stack images of Cav-1–/– endothelial monolayers pretreated with Rho inhibitor C3 transferase and stained for beta-catenin (green), F-actin (red) and nuclei (blue). Images were acquired using a Zeiss LSM 510 META confocal microscope equipped with c-Apochromat 63x/1.2 water correction objective.<br />

Beta-catenin distribution in Cav-1–/– endothelial monolayers pretreated with AP-CSD peptide: 12-bit projected image. <br />

FRET/CFP ratio images in Cav-1-/- endothelial monolayers. Pixel intensities of ratio images were scaled from 0 to 5 and color-coded.

FRET/CFP ratio images (RhoA activity) in Wt endothelial monolayers. Pixel intensities of ratio images were scaled from 0 to 5 and color-coded.

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