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.

Murine neural stem cell (mNSC) proliferation. A) mNSCs were treated with different concentration (0 nM, 1 nM, 10 nM, 100 nM) of methylphenidate (MPH). The percentage (% of control) of proliferated cells was determined 28 h after induction of neuronal proliferation and MPH treatment. The amount of proliferating cells was analyzed by counting the Bromodeoxyurdine (BrdU) positive cells in comparison to the total number of cells using the Mann–Whitney (U-Rang) Test; --- =p <0.05; n= 20; five independent experiments and four wells/slide of each concentration were evaluated. B) An example of an immunocytochemistry staining of BrdU in a control sample (no MPH treatment). The white arrow points to a BrdU positive cell (red) and the white arrow points to a BrdU negative cell (blue); 10x magnification.

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.