FIGURE 4.

Identification of FLNaABD lysine residues important for ASB2α-mediated degradation. A, CHO cells transfected with FLNaABD GFP, FLNaABD K33R/K42R/K43R GFP (K33/42/43R GFP), FLNaABD K135R GFP, and FLNaABD K42R/K43R/K135R GFP (K42/43/135R GFP) were fixed and stained for phalloidin (Phall). Scale bar, 10 μm. B, CHO cells from A were lysed and immunoblotted (IB) using anti-GFP antibody. C, CHO cells transiently expressing FLNaABD GFP, FLNaABD K33R/K42R/K43R GFP, FLNaABD K135R GFP, FLNaABD K42R/K43R/K135R GFP, and FLNaCH2 were transfected with either dsRed-ASB2α or dsRed-ASB2αΔS, and 48 h later the mean ± S.E. percentage of GFP-tagged protein remaining in dsRed-ASB2α-expressing cells was assessed as in Fig. 1 (see “Experimental Procedures” for details). Data are from at least three independent experiments.