Both the interaction and co-clustering of LCA with septins is impaired by the L428A/L429A mutation. (A) The L428A/L429A mutation decreases the amount of LCA that co-immunoprecipitates with septin-2. (B,C) The L428A/L429A mutation decreases colocalization of LCA with septin-2 (B) and septin-7 (C) both in non-differentiated and differentiated cells. The mutation also decreased accumulation of both septin-2 and septin-7 in LCA-positive regions. The coefficient of colocalization with septins (mean±s.d., n = 12) followed by the overlap coefficient (mean±s.d., n = 12) is indicated below the merged images. WT, wild-type GFP–LCA; NT, non-transfected cells; IP, immunoprecipitation; LL/AA, GFP–LCA-L428A/L429A mutation.