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. Author manuscript; available in PMC: 2009 Aug 1.
Published in final edited form as: J Immunol. 2008 Aug 1;181(3):2044–2055. doi: 10.4049/jimmunol.181.3.2044

Figure 6. PKD1 is required for CpG-B DNA-mediated activation of MAPKs and transcription factors.

Figure 6

A, Control luciferase-knockdown (L) or PKD1-knockdown (#1 and #2) RAW264.7 cells were transiently transfected with NF-κB-luciferase (left), AP-1-β-galactosidase (middle), or CREB-luciferase (right) reporter genes and then stimulated with medium (med), CpG-B DNA (6 μg/ml) or IFN-γ (10 ng/ml) for 12 h. Luciferase (NF-κB or CREB) or β-galactosidase (AP-1) activities in cell extracts were analyzed. Data represent the mean relative luciferase unit (RLU) ± SD of triplicates. B, Control luciferase-knockdown (L) or PKD1-knockdown (#1 and #2) RAW264.7 cells were stimulated with medium or CpG-B DNA for 1 h (NF-κB) or 4 h (AP-1). Cytoplasmic extracts and nuclear extracts were prepared. DNA-binding activities of transcription factor, NF-κB or AP-1, in equal amounts of nuclear extracts (3 μg/lane) were analyzed by EMSA (gel-shift) and degradation of IκBα and IκBβ in cytosolic extracts was detected by Western blot analysis (WB). C and D, Control luciferase-knockdown (L) or PKD1-knockdown (#1 and #2) RAW264.7 cells were stimulated with medium, CpG-B DNA or IFN-γ for 45 min. Phosphorylation status of MAPKs (JNK, p38, ERK), STAT1, and CREB was detected by Western blot analysis.