the species used to make the secondary antibody, plus 0.1% Tx-100 in PBS. Primary antibodies (detailed above) were incubated overnight at room temperature in the blocking buffer corresponding to the species of the secondary antibody. For immunofluorescent staining, the NSCs were washed with PBS and incubated with an Alexa 488 or a 635 fluorophor conjugated secondary antibody and counterstained with 4', 6-diamidino-2-phenylindole (DAPI- A-T-specific DNA stain at 350nm wavelength, Invitrogen). Staining of undifferentiated NSCs was done in a 1.5mL Eppendorf centrifuge tube following the procedures above.
