Enteropathogenic E. coli acts through WASP and Arp2/3 complex to form actin pedestals - PubMed (original) (raw)

Figure 1

WASP is recruited through its GTPase-binding domain (GBD) to EPEC pedestals. a–d, Images of HeLa cells exposed to EPEC. Cells were stained with DAPI, to identify EPEC (a), anti-WASP polyclonal Ab (b) or rhodamine–phalloidin, to visualize actin (c). Arrowheads denote endogenous WASP-like protein localized in pedestals. In the merged images d, p and t, EPEC are pseudocoloured blue, WASP green, and actin red. In h and l, EPEC are pseudocoloured red, WASP blue, and actin green. e–h, Images of cells expressing Flag (Fl)-WASP-WT and exposed to EPEC. In this panel and in i–t, cells were stained with DAPI (e, i, m, q), anti-Flag mAb (f, j, n, r), and rhodamine–phalloidin (g, k, o, s). Arrowheads denote Fl-WASP-WT protein localized in pedestals. i–l, Images of cells expressing high levels of WASP-ΔC. Note that EPEC on expressing cells have no actin pedestals associated with them (arrowheads). m–p, Images of cell expressing Fl-WASP-ΔGBD. Note that WASP-ΔGBD was in pedestals but was not enriched relative to the cytoplasm. q–t, Images of cell expressing low levels of Fl-WASP-GBD. Note that the GBD protein localizes to pedestals. Scale bar represents 4 μm in d, 5 μm in h, 10 μm in l, and 2 μm in p, t.