Figure 5
Figure 5. KSHV infection increases endothelial permeability in a Rac1-dependent manner. (A) HUVEC and KSHV-HUVEC were seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. A sample of medium from the lower chamber was taken after 6 hours, and the amount of FITC-dextran in the lower chamber was measured in a plate reader. These are representative graphs of triplicate assays (#P < .01). (B) HUVEC and KSHV-HUVEC were seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (40 kDa) was placed in the upper well. A sample of medium from the lower chamber was taken after 6 hours, and the amount of FITC-dextran in the lower chamber was measured in a plate reader. These are representative graphs of triplicate assays (*P < .05). (C) HUVEC and KSHV-HUVEC were transfected for 48 hours with control siRNA or siRNA targeting Rac1 and then seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. The amount of FITC-dextran in the lower chamber was measured in a plate reader 6 hours later. These are representative graphs of triplicate assays (#P < .01). (D) HUVEC and KSHV-HUVEC were infected with adenovirus containing WT VE-cadherin or a phosphorylation-resistant mutant of VE-cadherin (YY/FF), and then seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. The amount of FITC-dextran in the lower chamber was measured in a plate reader 6 hours later. These are representative graphs of triplicate assays (#P < .01). (E) KSHV-HUVEC display increased migratory potential. HUVEC and KSHV-HUVEC were seeded in 1 well of a 6-well dish for 24 hours. After wound formation (with a P10 pipette tip), cells were imaged at 0, 6, and 12 hours. (F) The percentage of the gap filled with migrated cells at 6 hours was normalized to the gap measured at 0 hours. The measurements were averaged from 10 different experiments. Error bars represent SD.

KSHV infection increases endothelial permeability in a Rac1-dependent manner. (A) HUVEC and KSHV-HUVEC were seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. A sample of medium from the lower chamber was taken after 6 hours, and the amount of FITC-dextran in the lower chamber was measured in a plate reader. These are representative graphs of triplicate assays (#P < .01). (B) HUVEC and KSHV-HUVEC were seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (40 kDa) was placed in the upper well. A sample of medium from the lower chamber was taken after 6 hours, and the amount of FITC-dextran in the lower chamber was measured in a plate reader. These are representative graphs of triplicate assays (*P < .05). (C) HUVEC and KSHV-HUVEC were transfected for 48 hours with control siRNA or siRNA targeting Rac1 and then seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. The amount of FITC-dextran in the lower chamber was measured in a plate reader 6 hours later. These are representative graphs of triplicate assays (#P < .01). (D) HUVEC and KSHV-HUVEC were infected with adenovirus containing WT VE-cadherin or a phosphorylation-resistant mutant of VE-cadherin (YY/FF), and then seeded in the upper well of a Transwell chamber and allowed to grow to confluence. FITC-dextran (10 kDa) was placed in the upper well. The amount of FITC-dextran in the lower chamber was measured in a plate reader 6 hours later. These are representative graphs of triplicate assays (#P < .01). (E) KSHV-HUVEC display increased migratory potential. HUVEC and KSHV-HUVEC were seeded in 1 well of a 6-well dish for 24 hours. After wound formation (with a P10 pipette tip), cells were imaged at 0, 6, and 12 hours. (F) The percentage of the gap filled with migrated cells at 6 hours was normalized to the gap measured at 0 hours. The measurements were averaged from 10 different experiments. Error bars represent SD.

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