EV71 2A protease without amino acids 146-149 still retained its protease activity but lost its transcriptional activity. (A) HeLa cells were transfected with vector only (lane 1) or with the plasmid encoding the C-terminus of VP1 and wild-type 2A (lane 2), or with the plasmid encoding the C-terminus of VP1 and 2A deleting amino acids 146-149 (lanes 3 and 4). After transfection, cell lysates were analyzed by Western blot using mouse anti-V5 tag monoclonal antibody. The thin arrow indicates the uncleaved protein (VP1-2A in lanes 2-4) while the thick arrow indicates the 2A protein (lanes 2-4). Erk2 protein served as a loading control. (B) Growth of yeasts either mock-transfected or transfected with plasmids encoding EV71 2A or 2A protein without amino acids 146-149 in YEPD medium, YEPD without tryptophan, or YEPD without tryptophan and histidine.