Figure 5.

Association of coronin 1 with the cytoskeleton. (A) Biochemical analysis of F-actin association of coronin 1. HEK293 cells transiently transfected with Cor1-HA, Cor1-Δ CC-HA, or Cor1-Δ400-416-HA were lysed in cytoskeleton isolation buffer containing 1% Triton X-100, and a detergent-insoluble fraction was obtained by low-speed centrifugation (see Materials and Methods). Subsequently, detergent-insoluble pellets and supernatants were analyzed by SDS-PAGE and immunoblotting by using an anti-coronin 1 antibody. (B) Schematic representation of coronin 1 deletion mutant Cor1-Δ400-416-HA. (C) Size exclusion chromatography analysis of cytosol from Cor1-Δ400-416-HA expressing HEK293 cells. Size exclusion chromatography was performed as described in Figure 3A and Materials and Methods.