Skip to main content
. 2017 Sep 29;4(5):ENEURO.0214-17.2017. doi: 10.1523/ENEURO.0214-17.2017

Figure 2.

Figure 2.

In vitro characterization of cultivated WT and Manf-/- NSCs. A, Immunofluorescent staining with MANF (green) antibody shows high expression in WT neurospheres, and no signal in Manf-/- neurospheres. B, There was no difference in the self-renewal assay in NSCs of Manf-/- and WT groups (p = 0.25, n = 4). Immunofluorescent staining with Nestin (green, C), DCX (red, E) antibodies and counterstained with DAPI (blue) in WT and Manf-/- NSCs. The percentage of Nestin-positive NSCs (D) and DCX-positive (F) neuronal precursor cells in WT and Manf-/- NSC cultures were analyzed by counting the ratio of Nestin- or DCX-positive cells to DAPI-positive nuclei, n = 3–5. NSCs were analyzed by Western blotting for Nestin, DCX, GFAP in WT and Manf-/- NSCs (G). Quantitative data from Western blottings (H) are presented as levels relative to GAPDH, n = 3–5. Numbers on the right show molecular weight as kDa. Cell viability did not differ between WT and Manf-/- NSCs (I). J, K, NSC proliferation assessed from BrdU-positive cells (red) as a ratio from DAPI-positive nuclei (blue) from NSC cultures, n = 5–6; scale bar: 50 µm.