M o m blocking kit

Post-fixed E10.5, E12.5, E15.5, E16.5 and P0 mouse brains were sectioned at 20 μm. Sections were fixed in 4% PFA in 0.1 m PB for 15 min, followed by blocking and permeabilization with 10% bovine serum albumin and 0.3% Triton X-100, and then finally treated with the M.O.M. blocking kit according to the manufacturer's instructions (Vector Laboratories, Burlingame, CA, USA). Mouse anti-Ki-67 (1:200, BD Bioscience), chick anti-GFP (1:400, Thermo Fisher Scientific), rabbit anti-GFP (1:400, Thermo Fisher Scientific), mouse (anti-phospho-histone H3 (pHH3)) (1:1000, Abcam, Cambridge, UK), rabbit anti-Pax6 (1:500, Merck Millipore, Billerica, MA, USA), rabbit anti-Tbr2 (1:500, Abcam), rabbit anti-PCDH19 (1:100, Merck), and mouse anti-SatB2 (1:200, Abcam) and anti-BrdU (1:200, Accurate Chemical and Scientific, Westbury, NY, USA) were used as primary antibodies. DAPI (1:1000, Dojindo) was used to counterstain the nuclei. For the proliferation and differentiation assays, sections with a similar anatomical EGFP distribution of EGFP expression were chosen selected for analysis, and a total of 3–5 sections were analyzed per embryo on a BX51 fluorescence microscope (Olympus) or FV-1200 laser-scanning confocal microscope (Olympus). The subventricular zone (SVZ) area was determined by DAPI staining, as described previously.^{19 (link)}