Murine neural stem cell (mNSC) differentiation into astrocytes. A) mNSCs were treated with different concentration (0 nM, 1 nM, 10 nM, 100 nM) of methylphenidate (MPH). The percentage (% control) of developed neurons was determined 4 days after treatment with MPH. The amount of astrocytes was analyzed by counting the glial fibrillary acidic protein (GFAP) positive cells in comparison to the total number of cells using the Mann–Whitney (U-Rang) Test; --- =p <0.05; n= 28; seven independent experiments and four wells/slide of each concentration were evaluated. B) An example of an immunocytochemistry staining of GFAP in a control sample (no MPH treatment). GFAP positive cell (red) and cell nucleus staining with Hoechst (blue); 40x magnification.