Figure 1.

EspG Interference of the WAVE Regulatory Complex

(A) Phagocytosis of pH-Rodo-labeled wild-type and ΔT3SS EPEC. Merged images show intracellular bacteria (red), extracellular bacteria (blue), and host cells (actin). Grayscale shown for clarity. Scale bars, 6 μm.

(B) Bar chart quantifying phagocytosis from experiment in (A).

(C) Phagocytosis of EPECΔ^T3SS (ΔT3SS) in THP1s transfected with Hem, Rac1, or Arf1 siRNA.

(D) Phagocytosis of EPECΔ^espG (ΔespG). Asterisks indicate a significant difference from control (black bars).

(E) Actin-based motility of PIP3-containing microspheres (depicted in cartoon) in cell-free extract containing a Rac1 inhibitor (+EHT1864) or EspG. Insets magnify actin-comet tails. Scale bars, 5 μm.

(F) WRC-dependent actin-based motility of PC:PI phospholipid bilayers microspheres co-anchored with Arf1^QL and Rac1^QL in cell-free extract containing fluorescent rhodamine-actin in the presence (+EspG) or absence (control) of recombinant EspG.

(G) Proteins recruited by PC:PI-coated microspheres alone (control) or co-anchored with Arf1^QL and Rac1^QL (colored circles) from extract (–/+ EspG) were analyzed by SDS-PAGE and Coomassie blue staining. Green arrows indicate cyfip, Hem, or actin. Orange arrows indicate absence of cyfip and Hem. Molecular weight markers in kilodaltons (left).

(H) Immunoblotting of samples from (G) with indicated antibodies (right).

In bar charts (B)–(D), error bars represent ± SEM. See also Figure S1.