FIG 2.

Screen of NleD multiple site-directed mutants for loss of proteolytic activity against JNK and/or p38. (A) Immunoblots of JNK and p38 cleavage in HEK293T cells transiently transfected with GFP-NleD or GFP-NleD multiple site-directed mutants. Cell lysates were probed with anti-JNK and anti-p38 antibodies. Anti-GFP antibody was used to show the expression of GFP-NleD and GFP-NleD variants. Arrows, cleavage products. Actin was used as a loading control. The immunoblots are representative of those from at least 3 independent experiments. UT, untransfected. (B) Schematic diagram mapping triple alanine replacement sites in NleD that alter JNK and/or p38 cleavage. Light gray arrowheads, cleavage of JNK but little or no cleavage of p38; white arrowheads, total cleavage of both JNK and p38. aa, amino acids.