Misu Regulates Stem Cell Fate Figure 7. Lack of Misu affects self-renewal of epidermal stem cells. (A,B) Colony forming efficiency of flow-sorted bulge stem cells (A) and 56103 or 104 whole epidermal cells isolated from wild-type (wt) and Misu 2/2 back skin (B). Error bars represent SD (n = 3). (C) Representative culture dishes from (B). (D) Confocal images of wild-type and Misu 2/2 primary keratinocytes in culture stained for Misu (green) and DAPI (blue). (E–J) Ki67 staining (E,H) and Haematoxylin and Eosin staining (F,G,I,J) of epidermal sections taken at them same regions of dorsal skin at indicated postnatal days obtained from anagen in Misu 2/2 and wild-type mice. (K–M) Percentages of male wild-type and Misu-depleted males in the first adult anagen (K), the second adult anagen (L), and the first anagen in K14MisuD/D and Misuf/f mice (M) at time points indicated. Detailed numbers of animals are summarized in Table S1. (N–U) Delayed expression of signalling molecules in growing hair follicles of Misu 2/2 mice compared to wild-type littermates at P25: Lef1 (green) (N,O), hair keratin 72 (green) and Dlx3 (red) (P,Q), phospho-Smad1/3/5 (green) (R,S) and hair keratin 31 (green) and Gata3 (red) (T,U). Nuclei are counter stained with DAPI (blue). Insert in (O) shows higher magnification of matrix cells. Arrows in (I) mark dermal papilla and arrows in (N–U) indicate matrix cells. Scale bars: 50 mm (D), 200 mm (E–J) and 100 mm (N–U). doi:10.1371/journal.pgen.1002403.g007 PLoS Genetics | www.plosgenetics.org 9 December 2011 | Volume 7 | Issue 12 | e1002403