As shown, outgrowths generated from the parental CDβ cells consisted of ductal and alveolar structures.
Figure 1 shows whole mounts and hematoxylin and eosin staining of outgrowths generated by the selected clones.
The outgrowths generated were positive for both luminal and basal markers and expressed the appropriate genotype (Additional file 9D,E).
PR expression was solely extranuclear in the outgrowths generated from all ductal, alveolar and multipotent progenitor clones (data not shown).
As described earlier regarding Figure 2, the outgrowths generated by transplantation of the ductal progenitor clone showed a normal ductal phenotype as well as an IDH phenotype.
Native mammary glands from pubertal mice and mammary outgrowths generated by CD24+ CD29hi MEC transplantation were mounted onto microscope slides and fixed in 10% formalin overnight.
Additionally, assessment of outgrowths generated from these blastocysts showed that the perturbing conditions of a simple medium had lasting effects on the gene expression of the outgrowths, with altered gene expression of Atrx and Nanog.
Unlike the previous report by Van Keymeulen and colleagues, the outgrowths generated in these experiments were not lineage-restricted since the engraftments contained both basal and luminal cells that expressed GFP (Additional file 8).
The expression of mammary-specific basal markers p63, CK5 and SMA, as well as luminal markers CK8, NKCC1 and ER, was examined using IF in outgrowths generated from the distinct progenitor clones.
