Stochastic 10-cell profiling of an MCF-10A clone expressing chimeric EGFR and Erbb2 receptors. Stochastic 10-cell profiling of an MCF-10A clone expressing chimeric EGFR and Erbb2 receptors

ErbB2 activation of MCF10A cells gives rise to a multiacinar phenotype that is incompletely penetrant. To identify candidate upstream regulators of the phenotype, an MCF10A-5E clone was subcloned to express chimeric receptors of human EGFR and rat Erbb2, and this B2B1 subclone was stochastically profiled (PMID: 20228812) after 24 hours of 3D culture with or without synthetic dimerization. Overall design: B2B1 cells were grown in 3D culture for six days, treated with or without synthetic heterodimerizer (AP21967) for 24 hours, and cryoembedded for frozen sectioning. 10-cell samples or pool-and-split controls were randomly collected by laser-capture microdissection for analysis of transcriptional regulatory heterogeneities by stochastic profiling (Janes et al., Nat Methods 7(4):311-7 [2010]).