Fig. 3: Lemd3 depletion results in the loss of the contractile phenotype in VSMCs in vivo.

a Schematic view of the generation of tamoxifen-inducible smooth muscle cell-specific Lemd3 knockout mice. LoxP sites are indicated by red triangles. b Representative western blot and quantification of protein expression in the aortas of 12-week-old Lemd3^WT mice and Lemd3^SMKO mice. GAPDH was used as an internal control. The data are presented as the relative fold changes compared with Lemd3^WT mice. n = 6 mice per group. c Representative images and quantification of immunohistochemical staining for ACTA2 and CNN1 in the thoracic aortas from 12-week-old Lemd3^WT mice and Lemd3^SMKO mice. Scale bar = 100 µm. n = 4 mice per group. d Contraction of aortic rings isolated from 12-week-old Lemd3^WT mice and Lemd3^SMKO mice in response to phenylephrine (Phe). n = 5 mice per group. e Left, Hematoxylin and eosin (H&E) staining of the carotid arteries of sham-operated and wire-injured Lemd3^WT mice and Lemd3^SMKO mice on Day 28 after surgery. The inner circles represent the neointimal areas, and the outer circles represent the medial and neointimal areas. Scale bar = 100 µm. Right, quantitative analysis of the intima areas, the media areas, intima to media ratio, and external elastic lamina (EEL) circumference in H&E-stained sections of the carotid arteries of wire-injured Lemd3^WT mice and Lemd3^SMKO mice at 28 days after surgery. n = 6 mice per group. f EdU staining of the carotid arteries of Lemd3^WT mice and Lemd3^SMKO mice at 28 days after wire injury. The white arrows mark the EdU-positive cells in the neointima and medial area. Mouse vascular elastic lamina (EL) exhibits green autofluorescence. Scale bar = 100 µm. n = 6 mice per group. HE hematoxylin and eosin, EEL external elastic lamina. Error bars indicate s.e.m. Statistical analysis was performed using two-sided unpaired Student’s t test with Welch’s correction for (b-d) and the Mann‒Whitney U test for (e) and (f).