Anti nestin

Brains were cut into 10‐µm‐thick cryosections. For immunofluorescence analyses, brain slices were washed three times in PBS (10 min each time). To detect BrdU incorporation, fixed brain slices were pre‐treated with 1 M HCl for 30 min at 37°C. The slices were then washed with 0.1 M borate buffer (pH 8.5) for 30 min (15 min each time) and with PBS for 30 min (10 min each time). Slices were fixed in 4% paraformaldehyde for 15 min at room temperature and washed three times in PBS (10 min each time). Next, the slices were blocked in a blocking solution (2% bovine serum albumin, 0.3% Triton X‐100 and 0.1% sodium azide) at room temperature for 2 h. The slices were then incubated with the primary antibodies (anti‐ARID1A, Sigma, HPA005456; anti‐Nestin, Aves Labs, NES; anti‐Tuj1, Biolegend, 801202; anti‐PAX6, Biolegend, 901301; anti‐BrdU, abcam, ab6326‐125; anti‐PHH3, Millipore, 09‐797; anti‐Ki67, Labvision, RM‐9106‐S1) diluted in the blocking solution at 4°C overnight and labelled using the appropriate secondary antibodies (Goat anti‐mouse 488, A11001; Goat anti‐rabbit 488, A11034; Goat anti‐rabbit 568, A11011; Goat anti‐rat 568, A11077; Goat anti‐chicken 488, A11039; Donkey anti‐goat 568, A11057) at room temperature for 2 h.