FIGURE 5.

Mint2 overexpression attenuates NGF-induced PC12 cell differentiation. A, Western blot showing Mint2 expression in DRG neurons but not in PC12 cells. B, PC12 cells were cotransfected with pEGFP-N2 together with pcDNA3 as control or with pcDNA3-Mint2. After 3 days with or without NGF, neurite outgrowth was assessed as an indicator of neuronal differentiation. Scale bar, 10 μm. C, quantification of neuronal differentiation of PC12 cells. Values represent the mean ± S.D., and the double asterisk indicates significant difference (p < 0.01, Student's t test). D, Western blotting was carried out to detect the levels of TrkA, p-TrkA, and p75 proteins in various groups. After NGF treatment, decreased levels of p-TrkA were detected in Mint2-overexpressing PC12 cells compared with those seen in wild type PC12 cells. E, coimmunoprecipitation (IP) analysis showing that the interaction between TrkA and Mint2 was not affected by NGF stimulation. IB, immunoblot.