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.
Smeets et al. BMC Immunology 2012 13:12 doi:10.1186/1471-2172-13-12