Extended Data Fig. 7: State assignments for HSC progeny.
From: Single-molecule imaging of transcription dynamics in somatic stem cells
a, Gating strategy. Left, removal of megakaryocytes occurs first. Right, cells with more than 10 copies of Gata1 are assigned to G1/2H, whereas cells with more than 150 copies of PU.1 are assigned to macrophage. b, Probability density distributions for PU.1 (left) and Gata2 (right) with overlaid fits for a two-component negative binomial distribution amongst cells after removing megakaryocytes, G1/2H, and macrophage . c, Bivariate distribution of the same cells. Contrary to the case in CMPs, the population of Gata2highPU.1high HSC progeny all had morphological characteristics similar to macrophage-like cells seen in GMP datasets, which also were Gata2highPU.1high (see Extended Data Fig. 3). As such, all cells for which PU.1 > 75 and < 150 were assigned to P1H. d, Probability distribution for Gata2 in the remaining cells, fit with a two-component negative binomial. e, A distribution such as that in d cannot be definitively separated into high and low components owing to overlap in the distributions; therefore, cells are assigned probabilistically during KCA to the G2H or LES state in order to correct for false transitions arising from uncertainty in the assignment. See Supplementary Discussion for more details on the rationale and implementation of probabilistic gating.
