Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System
Figure 5
Cleavage of HK on hantavirus-infected HUVEC.
HUVEC were plated and mock- infected or infected with HTNV or ANDV for 4 days. 50 nM of HK in buffer was added to the monolayer and incubated for 1 h at 37°C. The cells were incubated an additional 1 h with (A) 50 nM FXII and 50 nM PK in the absence or presence of 1 µM CTI or (B) 50 nM PK. To examine cleavage of HK, cells were lysed and prepared for immunoblotting. Full length, heavy, and light chain fragments of HK, hantavirus N, or GAPDH antibodies were used to detect protein. Levels of full length HK were analyzed by densitometery. (C) Samples were examined by immunoblotting to assess the levels of PRCP and HSP90 in mock-, HTNV-, and ANDV-infected HUVEC.