PMA/CD28-induced CCL1 production is not dependent on the Lck/Cn pathway. Jurkat T cells were stimulated using PMA/CD3 and PMA/CD28 in the presence of Lck (A420983), Cn (CsA) and PKC (AEB071) pathway inhibitors for 24 hours. Dose- response effects of the inhibitors were evaluated on the production of CCL1, after PMA/CD28 stimulation (A) and IL-2 after PMA/CD3 stimulation (B) in supernatant of the cell cultures. The data are representative for 3 independent experiments. C; Knock down of Lck and PKCθ resulted in a clear dose-dependent reduction of the protein (500, 100, 20 nM siRNA). 24 hour culture supernatants were collected after stimulation with PMA/CD28 and PMA/CD3 and the effect of knock down on respectively CCL1 and IL-2 was determined. Data of two independent experiments are presented as mean + SEM. Significance of differences are indicated by ** p < 0.01, *p < 0.05, # no difference using a one-way ANOVA with a Bonferroni's post-hoc test. N.b the values shown in this figure are log2 values no to be confused with the intensity values as shown in figure 3.