The effects on phospho-ERK1/2 and ERK1/2 by GTX65641-pro Progranulin protein in neuronal differentiated mouse P19 cells. Undifferentiated mouse P19 cells were induced to differentiated in 1microM retinoic acid (RA) in alpha-minimum essential medium (alphaMEM) containing 10% heat-treated fetal bovine serum on bacterial grade plates for 3~4 days to allow aggregates to form (generation of embryonic bodies). The aggregates were then plated on tissue culture grade plates in the absence of RA for 3~4days. To examine the induction of signal of phospho-ERK1/2 and ERK1/2, reactions were carried out at 37oC over 0, 5, 10, 30, 60, 120mins, respectively by adding the recombinant protein (500ng/ml) to the neuronal differentiated mouse P19 cells, which were maintained with serum starvation for 24hrs. Treatment with Progranulin protein (GTX65641-pro) was performed in lanes 1, 2, 3, 4, 5, and 6 over 0, 5, 10, 30, 60, 120mins, respectively. GAPDH was used as loading control for western blotting.