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. 2017 May 12;7(7):2033–2045. doi: 10.7150/thno.19824

Figure 3.

Figure 3

Effect of parkin and p21 on the expression of SNAP25 and BDNF. A, Neural stem cells isolated from Non-tg or PARK2 KO mice were harvested and western blotting was performed with p21, p53, p27, cyclinD1, pRb and Rb antibodies. Neural stem cells isolated from non-tg or PARK2 KO mice were differentiated for 5 days and then expression of SNAP25 or BDNF was visualized by Western blot analysis. B, Neural stem cells isolated from non-tg or PARK2 KO mice were transfected with p21 shRNA for 24hr, then differentiated, and then expression of SNAP25 or BDNF was visualized by Western blot analysis as described in the data. β-actin was internal control. Each band is representative for three experiments. C, Neural stem cells isolated from non-tg or PARK2 KO mice were transfected with p21 shRNA for 24hr, then differentiated into astrocytes and neuronal cells in vitro as described in materials and methods and then immunostained with GFAP or TUBBIII antibodies (upper panel). Number of neurite per cells and number of GFAP positive cells were quantified (lower panel). The data are expressed as the mean ± SD of three experiments. *P < 0.05 indicates significant difference from non-tg derived neural stem cells. #P < 0.05 indicates significant difference from PARK2 KO derived neural stem cells. D, Effect of p21 on the differentiation ability of parkin in vivo. Neural stem cells tranfected with parkin shRNA or p21 shRNA singly or together, and then injected in SVZ region of ICR mice. After 2 weeks, differentiated astrocyte or neuronal cells were stained with GFAP or TUBBIII (upper panel). Number of neurite per cells and number of GFAP positive cells were quantified (lower panel). The data are expressed as the mean ± SD of three experiments. *P< 0.05 indicates significant difference from control shRNA transfected neural stem cells. #P< 0.05 indicates significant difference from PARK2 shRNA transfected neural stem cells.