Cells were cultured in 8-well Lab-Tek Chamber Slides (Cat. No. 177402) for indicated time. Cells were fixed, permeabilized, blocked, and stained as described previously [28 (link)]. For indirect immunofluorescent cell staining, an anti-GAP43 monoclonal antibody and an Alexa Fluor 594-conjugated goat anti-mouse antibody were used to detect GAP43. Stained cells were imaged and analyzed using a Nikon Eclipse TE300 fluorescent microscope.
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