Anti sox 2

The slides containing brain organoid cryosections were washed twice with PBS (1x) and blocked with 0,1% TBST with 0.1% BSA diluted in PBS (1x). The following primary antibodies, diluted in 1% BSA, were incubated for 4 h at RT: anti-NESTIN (1:200, mouse—Millipore, Burlington, MA, USA), anti-SOX-2 (1:500, mouse—Biolegend, San Diego, CA, USA), anti-TBR1 (1:200, rabbit—Sigma Prestige, St. Louis, MO, USA), anti-β Tubulin III (1:1000, mouse—Promega, Madison, WI, USA), anti-DCX (1:1000, rabbit—Abcam, Cambridge, UK), anti-MAP-2 (1:500, mouse—Abcam), anti-GFAP (1:200, mouse—DAKO, Glostrup, Denmark), and anti-cleaved caspase-3 (1:100, rabbit—Millipore). The cryosections slides were washed twice with PBS (1X). The following secondary antibodies (1:1000, all from ThermoFisher) diluted in BSA (1%) were incubated for 2 h at RT: AlexaFluor 488 goat anti-mouse and AlexaFluor 594 goat anti-rabbit. The cryosections slides were washed twice with PBS (1x), followed by immunostaining with DAPI (1:1000) diluted in PBS (1x) for 30 min at RT. The cryosections slides were washed twice with PBS (1x), mounted with a mounting media, and stored at 4 °C.