Figure 5.

Effect of juvenile parkinson's disease related parkin mutation on p21 levels and neural stem cell differentiation. A, WT parkin in HEK293 cells decreased the steady-state level of p21, but that was blocked by parkin mutants (R275W and G430D). B, Effect of parkin mutation on the ubiquitination of p21. 293 cells were cotransfected with constructs as indicated. At 48 hours after transfection, cells were harvested and then immunoprecipitated with anti-p21. Ubiquitinated p21 was visualized by Western blot analysis using anti-ubiquitin. Each band is representative for three experiments. C, Neural stem cells were transfected with parkin wt, parkin R275W or parkin G430D, then differentiate into astrocytes and neuronal cells in vitro as described in materials and methods and then immunostained with GFAP or TUBBIII antibodies. The data are expressed as the mean ± SD of three experiments. *P< 0.05 indicates significant difference from control vector transfected group.