Fig. 2.

Dazl is required for restricting developmental potential of the germline in diverse strains of mice. (A) Developmental time course of germline using Dazl-tdTomato and Nanog:GFP reporters, detected by flow cytometry. At ∼12 ts (∼E11), most cells expressed only the Nanog:GFP reporter (green). As development proceeds, the proportions of cells expressing both fluorescent reporters (orange), or only the Dazl-tdTomato reporter (red), change. Numbers of embryos tested are listed in each column, and the fraction of cells expressing each reporter is shown as an average. (B) Flow cytometry for Nanog:GFP-positive cells of E15.5 control and Dazl-deficient ovaries of indicated strains. Autofluorescence in PE-Cy7 channel is shown on y axis. Red box indicates area in which Nanog:GFP-positive cells were counted. (C) Derivation of EG cell lines from control and B6.Dazl-deficient embryos. Cells were collected by fluorescence-activated cell sorting (FACS) at embryonic age indicated on x axis, and cultured under defined conditions. After 10 d, EG cell colonies were counted, and rate of EG cell derivation (per 100 EGFP-positive cells plated) was calculated. Number of embryos tested is listed in each column, mean + SD, *P value < 0.05, ** <0.01, *** <0.001, ns = not significant, using t test or Fisher’s exact test as appropriate.