The two NES-like leucine-rich domains in hA3G are nonfunctional. (A) Chicken muscle pyruvate kinase (CMPK) transfected into HeLa cells localized to the cytoplasm. (B) SV40 NLS added to the N-terminus of CMPK enables trafficking to the nucleus. hA3G with three leucine to alanine mutations (boxed in hA3G-NT and hA3G-CT from Fig. 1) in either the N or C-termini (C and E, respectively) or in both regions (G) localized in the cytoplasm when transfected into HeLa cells. An N-terminal SV40 NLS added to the hA3G mutants in C, E, and G, (D, F and H, respectively) were also localized in the cytoplasm when transfected into HeLa cells. The side-by-side panels for A–H are two representative fields of the same treatment group. DAPI staining indicates the position of the cell nuclei in each panel.