Fig. 2
Low-proliferative luminal-surface enlargement with epithelial rearrangement after E14.5. a Azan staining of developing trachea epithelium. b Representative 3D reconstructed images of an epithelial-cell cluster (25 cells). See also Supplementary Movie 2. c Apical-surface area. Numbers represent means. (n = 217, 253, and 189 in three embryos for E14.5, E16.5, and E18.5, respectively). center line, box limits, and whiskers represent mean, 25 and 75% confidence limits, and min and max values, respectively. d Proportion of luminal cells in total epithelial cells. Data represent means ± SEM (n ≥ 3) (n = 528, 732, and 1045 in three embryos for E14.5, E16.5, and E18.5, respectively). e Integration of quantitative values revealed that modest increases in cell numbers, apical enlargement, and apical emergence contributed to luminal-surface enlargement. f Optical image of tracheal epithelium in Nkx2.1CreERt2; LSLKrasG12D trachea and littermate control at E18.5. Whole tracheas were stained for CDH1 (green) and TOPRO-3 (red). g Gross morphology of Nkx2.1CreERt2; LSL-KrasG12D trachea and littermate control. Yellow arrows indicate trachea length. Tracheal tube length (h). External diameter (i). Data represent means ± SEM (n ≥ 3). P values (**P < 0.01, *P < 0.05) show the significance with the Tukey-Kramer method (c, d) or Student’s t test (h, i). NS not significant. Scale bar = 500 μm (g), 20 μm (f), 5 μm (b)
