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. Author manuscript; available in PMC: 2020 Apr 12.
Published in final edited form as: Circ Res. 2019 Apr 12;124(8):1253–1265. doi: 10.1161/CIRCRESAHA.118.314567

Figure 2. enSM22αgc promoter mediates SMC-specific gene expression.

Figure 2.

(A) Schematic design of SMC-specific PKCδ shRNA expressing vector. ZsGreen cDNA and PKCδ shRNA are driven by a chimeric promoter (enSM22αgc) that contains a smooth muscle-myosin heavy chain (SM-MHC) enhancer and a modified SM22α promoter whose G/C-rich cis-element is deleted. (B-D) enSM22αgc mediates gene expression specifically in SMCs (C), but not ECs (B) and adventitial fibroblasts (Adv) (D). Cells were infected with either the Lenti-CMV-ZsGreen-IRES-shPKCδ or the Lenti-enSM22αgc-ZsGreen-IRES-shPKCδ lentiviral particles. Bright-field (upper panel) and fluorescence (bottom panel) images of cells were taken 72h after transduction. Scale bar = 50 μm. (E) SMCs were transduced with lentivirus expressing non-targeting shRNA (shNT) or PKCδ shRNA (shPKCδ) under the indicated promoter. After 72h transduction, cells were harvested and the whole-cell lysates were subjected to immunoblot analysis.