FIGURE 3.

CH1 lysine to arginine substitution renders FLNaABD resistant to ASB2α-mediated degradation. A, CHO cells transfected with FLNaABD GFP, FLNaCH2 GFP, and FLNaABD Lys/Arg GFP were fixed and stained for phalloidin (Phall). Scale bar, 10 μm. B, CHO cells transfected with FLNaABD GFP and FLNaABD Lys/Arg GFP were lysed and immunoblotted (IB) using anti-GFP antibody. C, CHO cells transiently expressing FLNaABD GFP, FLNaCH2 GFP, and FLNaABD Lys/Arg GFP were transfected with either dsRed-ASB2α or dsRed-ASB2αΔS. 48 h after transfection, cells were detached and washed with PBS, and the GFP intensity of dsRed-expressing cells was measured. Bar chart depicts mean percentage of GFP-tagged protein remaining ± S.E. in dsRed-ASB2α-expressing cells normalized to levels in dsRed-ASB2αΔS-expressing cells (see “Experimental Procedures” for details). Data are from at least five independent experiments.