Fifty micrometers thick floating sections were incubated in blocking solution (PBS, 0.05% bovine serum albumin, 2% fetal bovine serum, 1% Triton X-100 and 0.1% saponin) for 30 min prior to addition of primary antibodies: rat anti-Ctip2 antibody (1:500; Abcam), rat anti-GFAP (1:2000, Invitrogen), chicken anti-GFP (1:1000, Abcam), rabbit anti-Iba1 (1:500, Wako), rabbit anti-LC3B (1:1000, Invitrogen), mouse anti-Parkin (1:200, Abcam), mouse anti-SQSTM1/p62 (1:200, Abcam), rabbit anti-Rab1A (1:200, Proteintech) and mouse anti-Satb2 (1:1000, Millipore) at 4°C overnight. After extensive washes, sections were incubated in appropriate secondary antibodies in blocking solution: Alexa Fluor 488 (1:1000, Invitrogen), and Cy3-conjugated (1:1000, Molecular Probes) for 2 h at room temperature. Antigen retrieval was performed for rat anti-Ctip2 antibody (1:500; Abcam) as previously described (22 (link)). Sections were counterstained with DAPI.