Inactive dCyaA-KP toxoid is endocytosed by a rapid macropinocytic pathway.

<p>(A) J774A.1 macrophages were incubated with BSA-Dyomics 547 (50 µg/ml) together with Alexa Fluor 488-labeled dCyaA or dCyaA-KP (1 µg/ml), respectively. After 10 and 30 minutes, the cells were washed, fixed and observed as above. (B) Co-localization of toxoids with BSA was quantified using ImageJ software as described in details in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002580#s4" target="_blank">Materials and Methods</a>. (C) J774A.1 cells were incubated for 30 minutes in HBSS with wortmannin (1 µg/ml), dynasore (40 µM) or chlorpromazine (5 µg/ml), before 5 µg/ml of Alexa Fluor 488-labeled dCyaA-KP was added and time course of toxoid uptake was followed by live cell imaging and (D) quantified as above (n = 3).