Figure 6.

Overexpressing NeuroD counteracts morphine-induced decrease in NeuroD activity. Primary cultures were treated with PBS, morphine (1 μm), morphine (1 μm) plus nd-vir, fentanyl (10 nm), fentanyl (10 nm) plus 190-vir, or fentanyl (10 nm) plus KN93 for 3 d. A, The amount of phosphorylated NeuroD (pNeuroD), total NeuroD, total CaMKIIα (CaM), phosphorylated CaMKIIα (pCaM), total ERK, and β-actin were determined as in Figure 1A. B, The levels of miR-190 and NeuroD (ND) mRNA were measured by real-time PCR. IP, Immunoprecipitation; IB, immunoblotting. C, The mRNA levels of DCX, ND4, Notch1, and Robo1 were determined by real-time PCR. A–C, Results were normalized against those in control samples. Error bars indicate SD. *p < 0.05.