FIG 2.

Adherens junction protein expression in T. gondii-infected HUVEC. (A) HUVEC were cultured to confluence on fibronectin-coated glass coverslips for 3 days and then either mock infected with fresh media or infected with T. gondii. At 18 hpi, the cells were fixed, permeabilized, and stained with antibodies specific to VE-cadherin and β-catenin and counterstained with DAPI. Bars, 20 μm. (B and C) The continuity of VE-cadherin and β-catenin signal at the cell periphery was quantified by examining immunofluorescent signal of VE-cadherin at the cell periphery and defining gaps of ≥2.5 μm as discontinuous signal. Data are presented as the means ± SEM from at least three independent experiments. Student’s t test was performed (***, P < 0.001). (D) HUVEC were cultured to confluence for 3 days and either mock infected with fresh media or infected with T. gondii. At 18 hpi, cells were lysed and analyzed by Western blotting using antibodies for total VE-cadherin, β-catenin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Representative blots from five independent experiments are shown. (E and F) Densitometry was performed on Western blots of VE-cadherin and β-catenin by normalizing band intensities to those of GAPDH. Data are presented as the means ± SEM. Combined data from five independent experiments are shown. Student’s t test was performed (*, P < 0.05).