Fig. 1.

HDAC9 upregulation is associated with TAA-related human genetic variation. a TGFβVs and SMCVs gene groups. b Heat map of mRNA expression of commonly dysregulated genes in siSMAD3 (HDAC9 fold change = 1.6) and siACTA2 (HDAC9 fold change = 2.0) treated human VSMCs. The scale bar indicates fold change Z-score. c QPCR validation of HDAC9 transcript induction in siSMAD3, siTGFB2, siACTA2 and siMYH11 treated cells. Four experimental replicates are plotted. d QPCR analysis of HDAC9 transcript in human VSMCs transfected with either empty vector, TGFR2^G357W, or ACTA2^R179H alleles. Four experimental replicates are plotted. e Quantitative western blot analysis of HDAC9 distribution in nuclear and cytoplasmic extracts demonstrates increased nuclear localization in the presence of TGFR2^G357W or ACTA2^R179H alleles. Three experimental replicates are quantified. f siHDAC9 treatment of VSMCs reverses MMP activity induced by expression of TGFR2^G357W or ACTA2^R179H alleles in human VSMCs as measured by in vitro gelatin zymography. g siHDAC9 treatment of VSMCs rescues inhibition of migratory activity induced by expression of TGFR2^G357W or ACTA2^R179H alleles in human VSMCs as assayed by wound healing assay. Serial photomicrographs of wound healing assay at 0 and 48 h are shown, VSMCs are false colored in red. Bar = 200 µM. h Quantitative western blot analysis of HDAC9 protein in nuclear extracts from human aortic tissue from TAA patients and non-aneurysmal controls. i Immunofluorescence staining demonstrates similar nuclear localization of HDAC9 protein in human TAA samples from diverse forms of TAA etiology. Aortic lumen is rightward facing. (20×, Bar = 50 µM; 100×, Bar = 10 µM). FTAAD, Familial Thoracic Aortic Aneurysms and Dissections; TAA, Thoracic Aortic Aneurysm. Bar graphs are presented as mean with error bars (±S.D.), Student’s T-test, *p < 0.05, **p < 0.01 vs. WT. Full-length western blots presented in Supplementary Fig. 8