Figure 10. Endocytosis of CyaA is not affected by cAMP.

(A) The capacity to translocate the AC domain across cytoplasmic membrane was assessed as the capacity to elevate cytosolic concentrations of cAMP. J774A.1 cells were incubated with CyaA or CyaA-KP-Alexa 488 for 30 minutes at 37°C and the amounts of accumulated cAMP were determined in cell lyzates. The shown curves are representative of two independent experiments performed in duplicates. (B) J774A.1 cells were incubated for 20 minutes at 37°C with 5 µg/ml of Alexa 488-labeled dCyaA-KP or CyaA-KP constructs, the protein uptake was analyzed by live cell imaging and (C) the numbers of endosomes in cytoplasm were quantified as above.