Figure 4. Prop1 regulates pituitary stem cells after birth.

(A) Number of colony-forming cells per pituitary after 10 days in culture (N = 3–6 pituitaries per stage). Two-way ANOVA *p<0.05 for Prop1^df/df vs aged matched Prop1^+/+ littermates, post hoc Fisher analysis. Plots denote the mean ± SEM and * indicates p<0.05 relative to control. (B and C) Pictures of the tissue culture plates showing colonies from Prop1 (left) and Pou1f1 (right) mutant pituitaries fixed and stained, show the different morphology of colonies from Prop1^df/df pituitaries (arrowheads). (C) Photos taken at higher magnification, 2x. (D) Time lapse microscopy shows the formation of a colony from a single stem cell captured 24 hr after the cells were plated and 4 days later. Colonies from Prop1 mutant pituitaries have an altered morphology since the colony formation starts around day 4. Scale bar 100 µm. See also Figure 4—figure supplement 1.

DOI: http://dx.doi.org/10.7554/eLife.14470.007

Figure 4—figure supplement 1. Pituitary stem-cell-derived colonies from Prop1 and Pou1f1 mutant pituitaries.

Immunostaining with specific antibodies performed on fixed colonies show expression of stem cell markers, SOX2, SOX9 and GFRa2 and CYCLIN D2. No differences were found between genotypes. Cell nuclei were stained with DAPI (blue).

DOI: http://dx.doi.org/10.7554/eLife.14470.008