Figure 6:
RNAscope assay for modSMC markers. A UMAP projection of 24-week Fbn1C1041G/+ and control SMC/modSMC clusters with expression plots for matrix metalloproteinase-2 (Mmp2) and osteoprotegerin (Tnfrsf11b) expression enriched in modSMC cluster. B Fraction of SMCs expressing Mmp2 and Tnfrsf11b in 4- and 24-week scRNAseq datasets. Cells with >1 transcript for denoted gene considered positive. *** denotes adjusted p <0.01 (Wilcoxon rank sum test). C Chromogenic amplified in-situ hybridization using sequence-specific probes for Tnfrsf11b (blue) and Mmp2 (red) in 4-week Fbn1C1041G/+ and control aortas identifies qualitatively increased Mmp2 expression with minimal Tnfrsf11b positivity in diseased samples. D 24-week aortic tissue RNAscope. Rare Mmp2-positive cells identified in littermate control animals, double-positive cells in Fbn1C1041G/+ samples (red arrows) represent modSMCs in tunica media. Adventitial fibroblasts display significant Mmp2 and rare Tnfrsf11b staining, confirming efficient hybridization and amplification in all samples. Images shown are representative of results for n=3 animals of each genotype.