Fig. 5

Dermal-derived Sfrp1 and Igfbp2 proteins induce the emergence of IFE Tbx3⁺ basal cells. a Immunofluoresence of α-SMA (green), vimentin (red) and Hoechst (blue) on ventral skin from virgin and 16 dpc mice. b Quantification of α-SMA⁺ cells in the vimentin⁺ cell population in the dermis. c, e FISH analysis of Sfrp1 and Igfbp2 mRNAs with anti-sense probes (green) in combination with immunofluoresence of α-SMA (magenta), vimentin (red) and Hoechst (blue) on ventral skin from 16 dpc mice. d, f Quantification of Sfrp1 ⁺ (d) or Igfbp2 ⁺ (f) cells in the α-SMA⁺/vimentin⁺ or α-SMA⁻/vimentin⁺ cell populations in the dermis. g–j Quantification of Ki67⁺ (g, i) and Tbx3⁺ (h, j) cells in IFE basal layers of ventral skin from 16 dpc mice (g, h) and virgin mice (i, j) injected with PBS, IWP2, Sfrp1 or Igfbp2. k, l Quantification of Ki67⁺ and Tbx3⁺ cells in IFE basal layers of ventral skin from 16 dpc mice injected with PBS, WAY316606-h or NBI31772. m, n Quantification of Ki67⁺ (m) and Tbx3⁺ (n) cells in basal layers of ventral skin from 16 dpc wild-type mice and 16 dpc Sfrp1 ^−/− mice interbred with wild-type males. o, p Quantification of the division pattern of Axin2-Cre-ER-labelled IFE basal cells (o) and K14-Cre-ER-labelled IFE basal cells (p) in ventral skin from virgin mice injected with PBS, IWP2, Sfrp1, Igfbp2 or Sfrp1 + Igfbp2. b, d, f–p Data represent the mean ± s.e.m. of averages of three independent experiments (n = 3). > 100 cells (b), > 140 cells (d), > 150 cells (f), > 500 cells (g–n), > 50 cells (o, p) were analysed to calculate the average in each experiment; *P < 0.05, **P < 0.01, ***P < 0.001, analysed by Dunnett’s multiple comparison test (b, g–l), the two-tailed t-test (d, f, m, n) or Tukey’s multiple comparison test (o, p). a, c, e White-dotted lines represent basement membranes. Experiments were repeated three times with three mice for representative images