Figure 3

Figure 3 CDKL1 is expressed in VSMCs of normal and diseased aortic tissue.

(A) Immunohistological analysis of CDKL1 expression in aortic wall tissue. Consecutive sections of formalin-fixed paraffin-embedded human aorta tissue from a control individual (control 1) and 2 patients with aortic aneurysms (aneurysm 1 and aneurysm 2) were immunostained for VSMC marker protein ACTA2 (smooth muscle actin), CDKL1 (antibody 1 [AB1; see Supplemental Data: Results]; 1:300), and the vascular endothelial marker CD31 using specific primary antibodies and horseradish peroxidase–conjugated (brown) secondary antibodies. Specimens were counterstained with hematoxylin (nuclei, blue). Representative pictures are shown. Note that CDKL1 is colocalizing with ACTA2-positive cells. Arrows indicate cells with stronger CDKL1 expression. Vv, vasa vasorum. Scale bar: 50 μm. (B) Reanalysis of combined publicly available human aortic single-cell RNA-Seq datasets (28, 29). Filtered and annotated data were merged and integrated using Harmony. Top left panel: Feature plot of CDKL1 expression in combined dataset. Each cell in the dataset is represented by a dot in the uniform manifold approximation and projection (UMAP) space. Cells with a CDKL1 expression above 0 are colored in blue; cells with no detected expression of CDKL1 are colored in yellow. Top right panel: UMAP representation of the annotation of the combined dataset. Annotation was copied and combined from original publications (28, 29). Bottom panel: Dot plot of marker genes confirming cell identity. Gene markers for specific cell types were selected from literature (25–27).