Extended Data Fig. 2: Comparative differentiation and long-term growth potential of E11.5 AGM cells based on PDGFRA, PDGFRB and Nestin-GFP expression.
(A) Dot plots showing % (percentage) conversion to marker-positive cells following differentiation induction of bulk cultured CD31-; PDGFRA+; Nestin-GFP− and CD31−; PDGFRA+; Nestin-GFP+ CFU-F cells (n = 3). (B) Dot plots showing % (percentage) conversion to marker-positive cells following differentiation induction of CD31−; PDGFRA+; Nestin-GFP− and CD31−; PDGFRA+; Nestin-GFP+ single cell derived large-, small- and micro- CFU-F colonies (n = 3). (C) (i) Isotype staining controls. (ii) Percentage of E11.5 Nestin-GFP+ AGM CD31–; PDGFRA+; Nestin-GFP– and CD31–; PDGFRA+; Nestin-GFP+ cells that also express PDGFRB. (D) Long-term growth of E11.5 AGM-derived CFU-Fs based on CD31, PDGFRA, Nestin-GFP and PDGFRB expression. (E) Confocal images showing vessel-like structures (left; longitudinal and right; cross section) lined by Nestin-GFP+ endothelial cells with surrounding PDGFRB+ pericytes. These images were taken from tissues harvested at 3 weeks after subcutaneous transplantation of a Matrigel plug loaded with PDGFRA+ Nestin-GFP–CD31–PDGFRB– FAC-sorted cells from E11.5 Nestin-GFP+ AGMs. Colony sizes: micro colonies (<2 mm, 2–24 cells), small colonies (2–4 mm, >25 cells) and large colonies (>4 mm, >100 cells). * p < 0.05, ** p < 0.01, *** p < 0.005; two-sided t-tests were used to compare staining (A and B), and a linear mixed model was used to compare the growth curves (D). Data were derived from biologically independent samples and experiments. Data represent mean ± SD. The precise p values are listed in the source data.
