ERK1/2 becomes activated independently of PKC and ROS on Don Q cells exposed to CpPLC.

<p>(A) Don Q cells were treated overnight with GF109203x (GF, 20 µM), 2-Aminoethoxydiphenyl borate (2APB, 70 µM) PD98059 (PD, 187 µM), Tiron (TIR, 2 mg/ml) or Helenalin (Hel, 1 µg/ml), previously to CpPLC (PLC) exposure (PC: positive control, NC: negative control, C: control of cells without CpPLC). Cells were collected and PKC activation was measured using Promega'sPepTag® Non-Radioactive Protein Kinase Assays, according to manufacturer instructions. Results are representative of two independent experiments. (B) Don Q cells were treated overnight with GF109203x (GF, 20 µM), PD98059 (PD, 187 µM), Tiron (TIR, 2 mg/ml) or BAY117085 (BAY, 0,004 µg/µl) or PMA (100 nM) as positive control (Y value = 1451±1095), then cells were lysed and immunoblot-ted against phospho-ERK. After stripping of the blot, total ERK1/2 was also evaluated. Densitometric analysis was determined using Image J software. Results are representative of five independent experiments (mean±SD) (*p<0,001 vs PLC).