title: GSM8128330: DMSO_P1_G06, scRRBS (multi-omics); Mus musculus; Bisulfite-Seq
library_construction_protocol: IdhR132H mouse glioma cells were cultured in the presence of 1 μM AG-881or vehicle (DMSO control) for > 2 weeks.  The treated cells were washed with cold PBS and then stained with 1 uM calcein AM (Life Technologies) and 0.33 uM TO-PRO-3 iodide (Life Technologies) for 30 min at 4°C. Sorting was performed with the FACS Aria Fusion Special Order System (Becton Dickinson) using 488 nm (calcein AM, 530/30 filter) and  640nm (TO-PRO-3, 670/14 filter). Side scatter (SSC) width versus forward scatter (FSC) area, and Trigger Pulse Width versus FSC criteria were used to discriminate doublets. Calcein AM positive and TO-PRO-3 negative live cells were sorted into 96 wells containing TCL buffer (QIAGEN) for the single-cell multi-omics experiment. The single-cell plates for the mouse glioma cells treaded by AG-881 or DMSO were profiled by the joint single-cell RRBS and whole transcriptome amplification (Smart-seq2) protocol. The library preparation and sequencing were performed as previously described (Chaligne et al, 2021). Single enzyme digestion (MspI) was used to enrich CpG dense regions in single-cell RRBS.