Murine neural stem cell (mNSC) differentiation into immature neurons. 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 immature neurons was analyzed by counting the neuron-specific class III beta-tubulin (Tuj 1) positive cells in comparison to the total number of cells by 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 Tuj 1 in a control sample (no MPH treatment). The white arrow points to a Tuj 1 positive cell (green) and the white arrow points to a cell nucleus staining with Hoechst (blue); 40x magnification.