Figure 4

Properties of neurites elongated from N2a-Rα cells. (A) Immunochemical staining for phosphorylated NF-H was used as an axonal marker in N2a-Rα and N2a-Vc cells. Cells cultured for two-days were stained using the anti-phosphorylated neurofilament 200kDa (NF-H) monoclonal antibody and the FITC-conjugated goat anti-mouse IgG antibody (upper two panels). The lower panels are bright-field images of the upper panels. It should be noted that the outgrowths on the N2a-Rα cells were clearly stained with the anti NF-H antibody (panel "N2a-Rα "). (B) The quantity of cdc42 mRNA was determined in N2a-Rα and N2a-Vc cells with real-time PCR using primers specific for cdc42. A high level of expression of cdc42 mRNA was observed in N2a-Rα compared with the N2a-Vc. The results are normalized to the expression level of cyclophilin mRNA and expressed relative to the value of N2a-Vc. Values are presented as the mean ± S. D. of four replicate determinations; **p < 0.01 and *p < 0.05