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. 2012 Aug 16;287(43):36147–36157. doi: 10.1074/jbc.M112.389676

FIGURE 5.

FIGURE 5.

FAK, ezrin, and PKCα are required for cryptococcal transmigration of HBMEC monolayer. A, HBMEC was transduced with FAK shRNA to knock down FAK expression. Western blotting with anti-FAK antibody showed the reduction of FAK in the cells with FAK shRNA but not in the scrambled shRNA cells. The transmigration assay was performed with the FAK knockdown cells. B, transmigration assay was done with normal HBMEC in the presence of FAK inhibitor-14. The number of transmigrated Cryptococcus cells was determined by plating the medium collected from the bottom compartment after 3 h of incubation. The result was normalized to show the relative transmigration rate compared with the untreated control. C, HBMEC was transfected with the wild type or DN T567A ezrin mutant construct. Transfection was verified by Western blotting with anti-VSVG antibody (bottom). The transmigration assay was performed with the ezrin transfected cells. D, a transmigration assay carried out with normal HBMEC in the presence of calphostin C, an inhibitor for PKC. *, p < 0.001.