Skip to main content
. Author manuscript; available in PMC: 2011 Mar 15.
Published in final edited form as: J Immunol. 2010 Feb 8;184(6):3145–3156. doi: 10.4049/jimmunol.0903718

Figure 3. Effect of pharmacological PKC/PKD inhibitor Gö6976 on SR-Ag-mediated activation of MAPKs and NF-κB and expression of cytokines and chemokines in vitro.

Figure 3

AMJ2-C11, MPRO-Neut, and MLE12 cells were pretreated with DMSO, Gö6976 (250 ng/ml), or Gö6983 (250 ng/ml) for 1 h and then stimulated with medium, SR-Ag (1 μg/ml for AMJ2-C11, 5 μg/ml for MPRO-Neut, 10 μg/ml for MLE12), PMA (10 ng/ml), PIC (100 μg/ml), CpG DNA (12 μg/ml for AMJ2-C11, 24 μg/ml for MPRO-Neut and MLE12), PGN (1 μg/ml for AMJ2-C11, 2 μg/ml for MPRO-Neut, 4 μg/ml for MLE12) or IFNγ (10 ng/ml) for 45 min (Panel A), 2 h (for IFNγ in Panel B), 4 h (Panel B), or 24 h (Panel C). Phosphorylation of pan-PKCs, PKD1, JNK, p38, ERK, and STAT1 and the presence of IκBα and IκBβ in cell lysates were detected by Western blot analysis (Panel A). Messenger RNA levels of the indicated cytokines and chemokines in cells were analyzed by RT-PCR (Panel B). Levels of the cytokines and chemokines in culture supernatants were analyzed by ELISA (Panel C). Data represent the mean concentration (pg/ml) ± SD of triplicates. Actin was used as a loading control. Experiments were done three times with similar results.