Immunolabeling for Cell Proliferation Markers

Every sixth section was immunolabeled for either BrdU, Ki67, or DCX as previously described (Wu and Hen, 2014 (link)) 2 weeks subsequent to the conclusion of behavioral procedures. Primary antibodies included: rat anti-BrdU (1:200, Serotec OBT0030; Kidlington, UK), rabbit anti-Ki67 (1:100, Vector VP-RM04; Burlingame, CA), and goat anti-DCX (1:500, SCBT sc-8066; Dallas, TX). Secondary antibodies included: Cy3 donkey anti-rat (1:800, Jackson 712-165-150; West Grove, PA), AlexaFluor 488 anti-rabbit (1:300, Molecular Probes A21206; Eugene, OR), and biotinylated donkey anti-goat (1:500, Jackson 705-065-147). Sections were imaged on an AxioObserver A.1 or Axiovert 200 (Zeiss; Oberkochen, Germany). All BrdU, Ki67, and DCX immunolabeled cells spanning the DG were quantified and multiplied by six.