Supplementary Figure 2: Active Wnt signaling in P7 and adult Lgr5–EGFP⁺ cells.

(Top) P7 Lgr5-KI/Axin-LacZ ovary section stained for LacZ activity (left) then co-immunostained with EGFP and K8 (right). Arrows point to Axin2⁺ Lgr5–EGFP^hi OSE cells. (Bottom) An adult Lgr5-KI/Axin-LacZ ovary section stained for LacZ activity (left) then co-immunostained with EGFP–HRP (middle). Black and white arrowheads point to Axin²⁺Lgr5–EGFP^hi and Axin2⁺Lgr5–EGFP^lo/neg OSE cells respectively. Neutral red counter-stain. qPCR with Axin2 (right) confirming expression in sorted Lgr5–EGFP^hi, Lgr5–EGFP^lo and Lgr5–EGFP^neg OSE cells, but not in OSE-stripped ovary cells. Histograms represent fold-change relative to Lgr5–EGFP^neg cells set as 1. Error bars indicate s.e.m. Gapdh was used as endogenous reference. Scale bars: 10 μm.