Effect of DADLE on phosphorylation of MAPK and Akt in PC12h cells: PC12h cells (0.8 million cells/60-mm dish) were differentiated with 100 ng/mL NGF for 72 h. The cells were re-fed once with 100 ng/mL NGF after 48 h of plating. After 72 h of differentiation, medium was removed from each dish and cells were re-fed with medium without NGF for 24 h. After 24 h of NGF deprivation, cells were treated with 10 nM or 10 μM DADLE for 0, 0.25, 0.5, 1, and 4 h. Cells were harvested and Western blotting was carried out for p-Akt (ser473), p-MAPK, total Akt and alpha tubulin as described in Materials and Methods. (A) DADLE (10 nM) induced phosphorylation of Akt and MAPK in PC12h cells; (B) Semi-quantification of phosphorylation of Akt and MAPK (10 nM DADLE); (C) DADLE (10 μM)-induced phosphorylation of Akt and MAPK; (D) Semi-quantification of phosphorylation of Akt and MAPK (10 μM DADLE). p-Akt was normalized to total Akt and p-MAPK to alpha tubulin. Relative O.D. was measured as described in Materials and Methods. Data are expressed as mean ± SEM of three independent experiments normalized to unit one for the 0 h treatment (control). * p < 0.05.