Figure 5
Clonal selection by homotypic cell aggregation in suspension with FSS. (a) Equal numbers of far-red-labeled GFP-expressing parental cells or clones W and H cells were mixed and suspended in culture medium followed by rotation for 3 h at 37 °C. (b) Representative images of cell aggregates composed of only GFP-expressing cells (upper), the mixture of GFP-expressing cells and far-red-labeled cells (middle), and only far-red-labeled cells (bottom). (c) Abundance ratio of each aggregate type. Error bars represent SEM of triplicate samples. *p < 0.005; †p < 0.01; §p < 0.0001 vs. GFP alone in each experimental group (two-tailed t tests). (d,e) Equal numbers of mCherry-expressing parental (Parental-mCherry) cells and GFP-expressing parental cells and clones W, EE, or H were mixed and suspended in culture medium followed by rotation for 22 h at 37 °C. Cells were centrifuged, plated in 6-well plates, and cultured for 6 days. Colonies of surviving cells with areas > 2500 μm2 were counted. (d) Representative images of colonies derived from the parental-mCherry and clone H cell mixture. (e) The number of GFP- or mCherry-positive colonies in each experimental group. Values are means ± SEM of triplicate samples. *p < 0.05 (vs. all other GFP-positive and mCherry-positive cells); n.s., vs. mCherry-positive cells in all other groups.
