Biochemical alterations of tight and adherens junction proteins induced by P falciparum sonicates in HDMECs. (A-C) HDMECs were cultured in 35-mm dishes until 3 days after confluence with or without parasite sonicates at 107 parasite equivalents/cm2 for 24 hours. Total cell lysates were fractionated into 4 subcellular fractions: cytosolic, membrane, nuclear, and cytoskeleton (from left to right). Fractions were analyzed by Western blot for the indicated tight and adherens junction proteins. (A) ZO-1 but not (B) VE-cadherin or (C) claudin-5 was redistributed from the cytoskeleton to other subcellular fractions. (D) HDMECs were treated as in panel A. Total protein level as determined by Western blot was reduced for claudin-5 but not for ZO-1 or VE-cadherin using β-actin as a loading control. Representative Western blot results and densitometry analysis of 3 independent experiments are shown. For densitometric analysis of subcellular fractions, each fraction was expressed as a percentage of the total densitometry measurements for all 4 fractions. *P < .05, **P < .01 compared with control values by Student paired t test. (A-D) Results are expressed as mean (± SD).
