Somatic cells suppress Wnt/β-catenin activity in PGCs. (A) Nuclear β-catenin (n-βcat, red), Axin2 (blue), and E-cadherin (magenta) immunofluorescence in E9.5 PGCs (green) cultured for 20 h in isolation (ISO) or with E9.5 tail somatic cells (+Soma) from tail somatic cells. Images are representative of mean values. Bar, 10 µm. (B) Quantification of n-βcat in cultured PGCs from A. n-βcat intensity for each PGC is normalized to the mean value of PGCs cultured in isolation in the same experiment. Each dot represents a single PGC; bars indicate the mean and boxes denote the middle 50% of data points. n = 3 experiments, 186–203 cells per condition; ***, P < 0.001 by Student’s t test. (C) Quantification of Axin2 in cultured PGCs from A. AXIN2 intensity for each PGC is normalized to the mean value of PGCs cultured in isolation in the same experiment. n = 4 experiments, 293–309 cells per condition; ***, P < 0.001 by Student’s t test. (D) Quantification of E-cadherin in cultured PGCs from A. E-CADHERIN intensity for each PGC is normalized to the mean value of PGCs cultured in isolation. n = 2 experiments, 102–114 cells per condition; ***, P < 0.001 by Student’s t test. (E) Percentile rank of individual cell n-βcat from B. (F) n-βcat (red) immunofluorescence in E9.5 PGCs (green) cultured for 20 h in isolation (ISO) or with conditioned media (+CM) from E9.5 tail somatic cells. Bar, 10 µm. Graph shows quantification of n-βcat in PGCs in these conditions. n = 4 experiments, 84–112 cells per condition; ***, P < 0.001 by Student’s t test.