Figure 2.

Depletion of Rac1, but not RhoA, reduces EA internalization by host cells. HeLa cells depleted of Rho-family GTPases were incubated with EA for 2 h, fixed with Bouin, and stained with Giemsa, and internalized parasites were quantified by optical microscopy. (A) Lysates of HeLa cells transduced by lentivirus containing shRNAi sequences and treated with puromycin for 2 weeks were evaluated for Rho GTPases expression by western blotting using actin or GAPDH as loading controls (detailed in Materials and Methods). (B) EA internalization was reduced approximately 35 and 76% in Rac1 depleted cell lineages (R30 and R75, respectively) when compared to the non-transduced group (-). (C) Cells depleted for RhoA had similar EA internalization when compared to the non-transduced groups (-). (D) EA invasion in cells transduced with scramble RNA sequences displayed no difference when compared to a non-transduced group (-). Results represent mean of at least three independent experiments in triplicate, and each dot represents one replicate (One-Way ANOVA, ^***P < 0.001, ^**P < 0.01, n ≥ 9).