Similar to what we found in the expression of pluripotent stem cell markers in iPS cells, we found no significant effects of ethanol on the expression of NPC markers, including Nestin, Pax6, and Sox2 (Fig. 2a and b). We then asked whether cell division is altered by alcohol exposure using 5-Ethynyl-2′-deoxyuridine (EdU) incorporation assays [23]. We found no statistically significant variation in the number of dividing cells in ethanol-exposed versus control cells (Fig. 2c and d). Likewise, ethanol had no effect on the number of self-renewing NPCs, as assessed by growth curve analysis (Fig. 2e).
