description: Protocols: We grew MCF10-2A TetOn-CENPA-FLAG-HA cells expressing either an empty control vector (p53-WT) or dominant-negative p53 (p53-DN) without Dox or with continuous exposure to Dox (10ng/ml, ++, chronic) for 69 days in parallel. Cells in the no Dox condition were split into two dishes at day 67 and Dox was added to one set on day 68 for 24h of CENP-A overexpression (10 ng/ml, +, acute) while the other remained without Dox (-, control). We prepared samples in singlet according to the 10x Genomics Sample Preparation Demonstrated Protocol. In brief, we harvested cells by trypsinization, resuspended in typical media and mixed thoroughly by pipette, passed cells through a 40µm cell strainer, and counted cells by Beckman Automated Cell Counter. We resuspended cells in 1x PBS + 0.04% BSA, mixed, centrifuged gently, aspired supernatant and repeated wash two times. We passed cells through another cell strainer (40µm), counted again and diluted to a final concentration between 700 and 1200 cells/µl. We then proceeded directly to GEM generation and barcoding using the Chromium Single-cell 3' Reagent Kits v2 protocol with a targeted cell recovery of 2000 cells per sample, followed by post GEM-RT cleanup and cDNA amplification. 3' Gene Expression Library Construction, according to the manufacturer's instructions.
attributes.tag: ENA first public
attributes.value: 2021-03-01
attributes.tag: ENA last update
attributes.value: 2020-12-09
attributes.tag: External Id
attributes.value: SAMEA7699198
attributes.tag: INSDC center alias
attributes.value: Nuclear Dynamics Unit UMR3664 CNRS/IC Institut Curie - Centre de Recherche
attributes.tag: INSDC center name
attributes.value: Nuclear Dynamics Unit UMR3664 CNRS/IC Institut Curie - Centre de Recherche
attributes.tag: INSDC first public
attributes.value: 2021-03-01T12:18:29Z
attributes.tag: INSDC last update
attributes.value: 2020-12-09T13:11:57Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9861:DD0d69
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: MCF-10-2A
attributes.tag: cell type
attributes.value: mammary gland epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: breast fibrocystic disease
attributes.tag: genotype
attributes.value: p53-DN TetOn-CENPA-FLAG-HA
attributes.tag: growth condition
attributes.value: no CENP-A induction
attributes.tag: organism part
attributes.value: mammary gland
attributes.tag: post analysis well quality
attributes.value: pass
attributes.tag: sample name
attributes.value: E-MTAB-9861:DD0d69
attributes.tag: single cell well quality
attributes.value: OK