Fig. 1. WAVE2 is localized at membrane ruffles and phagocytic cups in Vero cells. (A) Immunoblots of Vero cell lysates using anti-pan WAVE and anti-N-WASP antibodies (upper panel), and anti-WAVE1, anti-WAVE2 and anti-pan-WAVE antibodies (lower panel). N-WASP is detected as a protein of 65 kDa and WAVE at 80 kDa and 90 kDa, the latter reflecting phosphorylated WAVE. Actin was detected as a loading control. (B) Colocalisation of WAVE and actin filaments in areas of ruffling (indicated by arrows) in Vero cells stimulated with 4.5 nM InlB. N-WASP is not localized at InlB-induced membrane ruffles. Cells were stained with anti-pan WAVE or anti-N-WASP antibodies and phalloidin-488. (C) Recruitment of WAVE but not N-WASP to bacteria-induced phagocytic cups in Vero cells. The accumulation of WAVE and actin filaments is indicated by arrows. Cells were stained with anti-pan WAVE or anti-N-WASP antibody (red), anti-InlA to detect bacteria (blue) and phalloidin-488 (green). Bar, 5 µm (B) and 2 µm (C).