Fig. 1
Kcnj13T38C/T38C mice exhibit tracheal tube formation defects. a Workflow for ENU screen in mouse to identify mutants with respiratory organ formation defects. b Representative gross morphology of P0 WT (n = 12) and mutants (n = 12). c Quantification of P0 WT (n = 12) and mutant (n = 12) respiratory rates. d Representative image of ventral view of whole-mount tracheas stained with alcian blue from P0 WT (n = 35) and mutants (n = 35). Double-sided arrows indicate tracheal tube length. Arrows point to tracheal cartilage rings. e Quantification of P0 WT (10) and mutant (10) tracheal tube length. f Representative images of transverse sections of tracheas stained with alcian blue and nuclear fast red from P0 WT (n = 9) and mutants (n = 9). g Quantification of P0 WT (n = 9) and mutant (9) tracheal lumen area. h Representative images of dorsal views of whole-mount tracheas stained for αSMA (red) from P0 WT (n = 10) and mutants (n = 10). i Representative images of lung tissue sections stained with hematoxylin and eosin from P0 WT (n = 8) and mutants (n = 8). j Quantification of P0 WT (n = 8) and mutant (8) terminal airspaces in the lungs. k Whole-exome sequencing of P0 WT (n = 2) and mutant (n = 2) genomic DNA. Green indicates the WT nucleotide A. Light brown indicates the mutant nucleotide G. l Sequence of WT and mutant genomic DNA around the lesion. DNA sequence fluorogram shows CTG for leucine in WT (n = 119) (upper panel) and CCG for proline in mutants (n = 105) (lower panel). Arrows point to the mutation site. m Representative images of ventral views of whole-mount tracheas stained with alcian blue from P0 WT (n = 17) and Kcnj13Del-E2&E3/T38C double heterozygous animals (n = 17). Double-sided arrows indicate tracheal tube length. Arrows point to tracheal cartilage rings. Scale bars: 1000 μm (d, m), 500 μm (h), 100 μm (e, i). *P < 0.05; **P < 0.01; unpaired Student’s t-test, mean ± s.d. WT: wild-type, MT: mutant, A: anterior, P: posterior, D: dorsal, V: ventral, L: left, R: right, Cart: cartilage, SM: smooth muscle
