Figure 9. Modulation of actin dynamics by EhAK1.

(A) Effect of EhAK1 on F-actin content of E. histolytica cells. E. histolytica cells (2×10⁵) containing indicated constructs were grown in presence or absence of 30µg/ml tet or G418 for 48 h. Amount of F-actin was measured using TRITC-phalloidin as described in “Materials and Methods”. For phagocytosis, amoebic cells were incubated with RBCs for 10 min at 37°C. (B) Effect of EhAK1 on actin polymerization. Rabbit muscle actin (10∶1 unlabelled and pyrene-labelled actin) was polymerized with and without indicated proteins as described in “Materials and Methods”. The increase in fluorescence of pyrene was observed at 407 nm. (C) Critical concentration was measured as described in “Materials and Methods”. The steady state pyrene fluorescence was recorded after incubating actin as above at the indicated concentration for 16 h at 25°C in the presence of EhAK1 and K85A. *p-value≤0.05, **p-value≤0.005, ***p-value≤0.0005.