FIG 5.
Analysis of various NleDR203 site-directed mutants for p38 cleavage and their effect on AP-1 activation. (A) Immunoblots of JNK and p38 cleavage in HEK293T cells transiently transfected with GFP-NleD or GFP-NleD single 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) Fold increase in AP-1-dependent luciferase activity in HEK293T cells expressing GFP or GFP-NleD variants unstimulated or stimulated with PMA for 6 h. Results are the means ± SEMs from at least three independent experiments carried out in duplicate. *, a result significantly different from the results obtained with GFP-NleD only stimulated with PMA (P < 0.05, by an unpaired, two-tailed t test).