Multiome snRNA + snATAC on MCF10-2A cells to study how high CENP-A levels promote EMT
Multiome snRNA + snATAC on MCF10-2A cells to study how high CENP-A levels promote EMT
MCF-10-2A TetOn-CENPA-FLAG-HA cells stably expressing a dominant-negative p53 vector (p53-DN) were cultured for 24 days in the absence (no Dox) or presence of Dox (10 ng/mL) to induce basal or high CENP-A expression, respectively. Culture medium (+/- Dox) was renewed every 2-3 days. We harvested cells at days 10 and 24 post-induction and prepared 4 million cells/mL in ice-cold cryopreservation medium (50% fetal bovine serum, 40% growth medium and 10% dimethyl sulfoxide). Single-nuclei suspensions prepared using 10X Nuclei Isolation protocols. Library construction by Active Motif services using the 10x Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent kit. Steps included nuclei transposition, generation of Gel Beads in Emulsion (GEMs) and 10X barcoding, cleanup, pre-amplification of ATAC DNA and cDNA, amplification with P5 and P7 adapters, Illumina sequencing.