Cells were fixed using 4% paraformaldehyde (Sigma). Cells were permeabilized by incubating with 0.5% Triton-X for 10 min. Antigen retrieval for brdU staining was performed using beta-catenin pathway 100 units/mL of DNase I (Worthington, Lakewood, NJ) supplemented with 1 mM MgCl2. Cells

were blocked for 30 min in 3% donkey serum, 0.2% Triton-X, diluted in TBS. Primary antibodies were diluted in blocking buffer and were incubated overnight at 4°C. Secondary antibodies were incubated for 1 hr at room temperature in the dark. All cells were counterstained with DAPI (1:5000 diluted in TBS) and coverslips were mounted using ProLong Antifade (Invitrogen). Three biological replicates were performed per experiment. Six high-power fields were counted per coverslip, except in relation to FZD2 manipulation experiments, in which four high-power fields were counted per coverslip. Dual GFP/luciferase reporters were purchased from System Biosciences (Mountain View, CA) for TCF/LEF, AP1, cJun, and NFAT. Lentivirus was produced

to infect proliferating NHNP cells already harboring targeting and nontargeting Venetoclax cost GRN hairpins, producing eight different transduced lines. Six biological replicates of each line were induced for 1 week with doxycycline and compared to its uninduced counterpart. GFP fluorescence was assessed using

a Plate Reader (BioTek) to determine relative not activity for each reporter in each cell type. Progranulin knockout mice were created as reported previously (Kao et al., 2011). The mice used for the microarray study were backcrossed into the C57BL/6 strain twice. This work was supported by the Consortium for Frontotemporal Dementia (CFR). This work was supported by NIA 5R01AG026938 (D.H.G.), by NIH/NINDS Neurobehavioral Genetics Training Grant 5T32NS048004-05 (E.Y.R.), the John Douglas French Alzheimer’s Foundation and NIMH K08MH74362 (E.M.W.), and by NIH AG034793 (L.H.M.). We are also grateful to Jeremy Miller for critical reading of this manuscript and we would like to thank Lauren Kawaguchi for her expertise as laboratory manager. “
“Damage to the adult mammalian CNS, in stroke or in spinal cord injury, remains devastatingly untreatable.