Fig. 1

Crossbreeding was conducted between male MyoD-cre/Aco2 flox/ + mice and female WT/Aco2 flox/flox mice, and fetuses were sampled from the parent mice on gestational day 18.5. (A): Photographs of extracted fetuses. Samples 2–5 were obtained from Ctrl mice, and samples 1 and 6 were obtained from cKO mice. (B): Photographs of the diaphragm obtained from the fetuses shown in A. Samples 2 and 3 are from Ctrl mice, and sample 1 is from a cKO mouse. (C) Expression of Aco2 in diaphragm muscle fibers and its complete loss in cKO mice. Fluorescent immunostaining was performed on cross sections of diaphragm tissue from Ctrl and cKO mice using Aco2 (green), Laminin α2 (red), and DAPI (blue). Scale bars represent 50 µm. (D) β-galactosidase staining of diaphragm sections from Ctrl and cKO mice at E18.5. Scale bars represent 100 µm.