The effect of PAR activation on the kinetics of ERK1/2, p38 and Akt phosphorylation. HOKs were stimulated with thrombin (10 U/ml) or trypsin (10 nM) for 0-90 min. (a-b) Phosphorylation of ERK1/2 and p38 was quantified in cell lysates using ELISA. Data are given as percent of unstimulated cells and as means ± S.E.M from two different donors. (*p < 0.05 compared to unstimulated control at 0 min). (c-d) The Western blots analysis of whole cell lysates from cells stimulated with thrombin (10 U/ml) or trypsin (10 nM) at indicated times. Cell lysates were analyzed by immnoblotting with specific antibodies to phospho-ERK1/2, total ERK1/2, phospho-p38, total p38, phospho-Akt and total Akt. GAPDH was served as a control to verify equal loading and transfer of samples.