Fig. 2

EPE peptide treatment inhibits ERK1/2 nuclear translocation in COS7 cell line. COS7 cells were grown in DMEM supplemented with 10% FBS and then serum starved (16 h, 1% FBS), pretreated either with EPE (10mM, 2 h) or left untreated as control (1% FBS). Cells were then either stimulated with 12-O-Tetradecanoylphorbol-13-acetate TPA; (200nM, 15 min) or left untreated (NT) as control. A: amino acid sequence of synthesized inhibitory peptide, composed of TAT as a cell penetrating motif, HA-tag and EPE sequence. B: Inhibition of ERK1 nuclear translocation is presented as the ratio between nuclear ERK1 levels and cytosolic ERK1 levels. Results were normalized to unstimulated cells (N ≥ 3, p = 0.0171). C: Inhibition of ERK2 nuclear translocation is presented as the ratio between nuclear ERK2 levels and cytosolic ERK2 levels. Results were normalized to unstimulated cells (N ≥ 3, p = 0.0157). D: Inhibition of the nuclear translocation of ERK1/2 is expressed as the ratio between nuclear ERK1/2 levels and cytosolic ERK1/2 levels. Results were normalized to unstimulated cells (N ≥ 6, p = 0.0006) E: Representative Immunoblots of COS7 cells treated with EPE (10μM, 2 h) with or without TPA stimulation (100/200nM). All data are presented as mean ± SEM