GSM7439878: JCB-5 E12.5 GFP- GFP; Mus musculus; OTHER
The hindbrains of 28 E12.5 and 24 E14.5 Atoh1GFP/GFP embryos were dissected  in ice-cold HEBG media (0.8X B27, 0.25X GlutaMAX in Hibernate E media). Tissues were incubated with Worthington Papain solution at 37°C for 30 min at 800 rpm. At the end of incubation, the samples were transferred to 15 mL falcon tube, followed by gentle trituration with serologic pipette. The cell pellets were collected by centrifuge at 200 rcf for 3 min at 4°C, washed and resuspended in ice-cold sorting buffer (PBS with 0.05% fetal bovine serum). The single-cell resuspension was loaded to 30 µm cell drainer to remove debris, followed by DAPI staining at RT for 5 min. GFP+DAPI- and GFP-DAPI- cells were sorted by Sony SH800S. CUT&RUN was then performed on 200,000 sorted cells with an anti-GFP antibody and IgG antibody for each sample. Following protocols.IO (dx.doi.org/10.17504/protocols.io.bagaibse) using NEB Next Ultra II DNA Library Prep Kit (E7645S) and NEB Unique Combinatorial Dual index kit (E6442S) Sequencing at GENEWIZ using Illumina Novaseq S2 lane with 150bp PE reads