Mouse anti brdu clone bu20a

As described previously^{11 (link)}, we cloned TNFR1 cDNA (Sigma) into a synapsin promoter-driven lentiviral expression system, and cloned TNFR1 or TNFR2 shRNA (Sigma) into a lentiviral shRNA system. Lentiviruses were generated and amplified using HEK 293T cells and then purified as previously described^{11 (link)}. Brain histology was analyzed using brain sections and immunostaining. Briefly, mice under anesthesia were transcardially perfused with 4% PFA and brains were removed, fixed, and post-fixed in 4% PFA, and infiltrated with 20–30% sucrose. Tissue sections were cut according to 20 um thickness, blocked, penetrated with 0.2% Triton-X 100, treated with rabbit anti-TNFR1 (polyclonal, Abcam, 1:1,000), mouse anti-TNFR2 (clone L-20, Santa Cuz, 1:1,000), or mouse anti-BrdU (clone Bu20a, Cell signaling, 1:1000) primary antibody, and subsequently reacted with a fluorescent secondary antibody (Invitrogen, 1:1,000). BrdU staining: mice were injected with BrdU (Invitrogen) at the dose of 30 mg per kg of body weight for 2 h before these animals were perfused. Naive IgG (clone DA1E, Cell signaling) of appropriate species were used as negative controls. DAPI staining was used to reveal all cells in the sections. Images were captured using confocal microscopy.