BrdU Labeling and Quantification

Animals were injected with 50 mg/kg BrdU on 3 consecutive days and sacrificed after the indicated time. Sections were incubated with 2 M HCl in PBST (PBS+0.3% Triton) for 25 min at 37°C for denaturation of the DNA, neutralized with 0.1 M sodium tetraborate (pH 8.5) for 7 min at room temperature and stained with an anti-BrdU antibody (AbDSerotec). On day 107–109 at 12 a.m., the mice received a BrdU injection and at day 123, the animals were sacrificed. The brains were dissected and analyzed (Hillje et al., 2013 (link)). For quantification of BrdU+ cells in the DG, two sections each of 5 wt and 5 TRIM32 ko brains were analyzed, for quantification of BrdU+ cells in the OB, two sections of 4 wt and 4 TRIM32 ko mice were used. In each case, the mean of BrdU+ cells in TRIM32 ko tissue was normalized to the mean of BrdU+ cells in sections of wt brains.