A and B, RAW264.7 cells pretreated with DMSO or Src PTK inhibitors (PP1 or SU6656) for 30 min (A), or that constitutively over-express empty vector or mCsk (B) were stimulated with medium or CpG-B DNA (12 μg/ml) for 45 min. C–E, RAW264.7 cells were pretreated with DMSO, PI3K inhibitor (LY294002), PLC-γ inhibitor (U73122), PKC inhibitors (Gö6850, Gö6976 or Gö6983 at 125 ng/ml), or EGTA for 30 min and then stimulated with medium, CpG-B DNA, or PMA for 45 min. Phosphorylation status of PKD1 and PTKs was detected by Western blot using a phospho-PKD specific antibody and phospho-tyrosine specific antibody (4G10), respectively. med: medium, SU: SU6656, LY: LY294002, U73: U73122.