FIGURE 4.
ELMO recruits ACF7 to the membrane. A, down-regulation of ELMO2 expression by siRNA was verified by Western blotting. B–D, ELMO2 is required for ACF7 recruitment to the membrane: MDA-MB-231 cells transfected with HA-ACF7 (B), Myc-ELMO1 and HA-ACF7 (C), and Myc-ELMO1PxxP and HA-ACF7 plasmids (D) together with either a siRNA targeting ELMO2 or a nonspecific siRNA were serum-starved. The cells were plated on fibronectin-coated chambers for 2 h and stained for ELMO1 (anti-ELMO1 for endogenous or anti-Myc for exogenous ELMO1), ACF7 (anti-HA) and DAPI. The arrows represent recruitment of ACF7 at the membrane. TCL, total cell lysate.