Figure 6.

FACS isolation of Sox9^EGFPpopulations. (A) FACS isolation strategy for Sox9^EGFP populations demonstrating that Sox9^EGFP is divided into 4 populations by flow analysis. (B) FACS-isolated cells from Sox9^EGFP populations exhibit increasing EGFP intensity (scale bar = 100 μm). (C) GFP^neg cells are the most abundant population present in single cell preps after exclusion of CD31/CD45/Annexin V/7-AAD⁺ cells, and (D) GFP^high cells are the most abundant of GFP⁺ populations (∗ indicates significance, P < .05, one-way analysis of variance and Tukey test). (E) Cell area measurements demonstrate that isolated GFP^high cells are significantly smaller than GFP^low cells (n = 150 GFP^low, 150 GFP^high; ∗ indicates P < .001, unpaired t test).