Fig. 2

Tbx3 is required for proliferation of IFE basal cells and abdominal skin expansion during pregnancy. a qPCR analysis of Tbx3 gene expression in dorsal and ventral skin from virgin and pregnant (8–16 dpc) mice. Data represent the mean ± s.e.m. (n = 3 mice); *P < 0.05, **P < 0.01, analysed by Dunnett’s multiple comparison test. b Immunofluoresence of Tbx3 (green) and Hoechst (blue) on ventral skin from virgin and 16 dpc mice. c Quantification of Tbx3⁺ cells in IFE basal layers. d, f Immunofluoresence of Tbx3 (d) or Ki67 (f) (green) and Hoechst (blue) on tamoxifen-treated dorsal and ventral skin of 16 dpc control (Tbx3 ^flox/flox /WT) and Tbx3 cKO (Tbx3 ^flox/flox /K14-CreER) mice interbred with genetically unaltered males. e, g Quantification of Tbx3⁺ cells (e) and Ki67⁺ cells (g) in IFE basal layers of dorsal and ventral skin from 16 dpc mice. h Representative images of 16 dpc control and Tbx3 cKO mice. An arrow indicates a swollen flank. i Representative images of razor-cut abdominal skin of 16 dpc control and Tbx3 cKO mice. The skin was laterally cut with a razor at 1.5 cm in length. Arrows indicate the wound width. j Quantification of the wound width. k Thickness of keratin 10⁺ layers of ventral skin from 16 dpc control and Tbx3 cKO mice. l IFE basal cell density in ventral skin from 16 dpc control and Tbx3 cKO mice. m Immunofluoresence of α18-catenin (green) and Hoechst (blue) on ventral skin from 16 dpc control and Tbx3 cKO mice. c, g Data represent the mean ± s.e.m. of averages of three independent experiments (n = 3). > 500 cells (c), > 220 cells (e), > 200 cells (g) were analysed to calculate the average in each experiment; **P < 0.01, ***P < 0.001, analysed by the two-tailed t-test (c), Dunnett’s multiple comparison test (e) or Tukey’s multiple comparison test (g). j–l Data represent the mean ± s.e.m. (n = 3 mice); *P < 0.05, **P < 0.01, analysed by the two-tailed t-test. b, m White-dotted lines indicate basement membranes. Experiments were repeated three times with three mice for representative images in (b, d, f, h, i, m)