Figure 3.

High GATA6 Expression Results in a Reduced Cloning Efficiency

(A) Percentage cloning efficiency of each cell fraction (3+/6−, 3+/6L, 3+/6H, and 3−/6+) using OCT4 (left) and SOX2 (right) as markers for the stem cell state. Sorted fractions were plated as single cells at clonogenic density in KO/SR and MEF conditions. Cloning efficiency was calculated by dividing the number of OCT4-positive (left) or SOX2-positive (right) colonies by starting seed density. Error bars represent SD of three biological experiments. Student’s t test was used to determine significance (OCT4 graph: 3+/6− to 3+/6H *p = 0.0017, 3+/6− to 3−/6+ ^∗∗p = 0.0001, SOX2 graph; 3+/6− to 3−/6H ^∗p = 0.0019, 3+/6− to 3−/6+ ^∗p = 0.0002).

(B) Proportion of OCT4-positive (OCT4[+]) cells in OCT4-positive colonies derived from single cells from fractions 3+/6−, 3+/6L, 3+/6H, and 3−/6+. Positive colonies include one or more OCT4(+) cells. Counts are shown as bar plots (blue) with superimposed estimated nonparametric distribution (red).

(C) Kullback-Leibler symmetric divergence between OCT4-associated distributions shown in (B). This measure increases with reduced similarity between distributions; zero indicates identical distributions.

(D) Proportion of SOX2(+) (SOX2-positive) cells in SOX2-positive colonies derived from single cells from fractions 3+/6−, 3+/6L, 3+/6H, and 3−/6+. Positive colonies include one or more SOX2(+) cells. Counts are shown as bar plots (blue) with superimposed estimated nonparametric distribution (red).

(E) Kullback-Leibler symmetric divergence between SOX2-associated distributions shown in (D).