Fig. 1

RGS5 ablation promotes VSMC inflammatory response and vascular remodeling. (A) Western blot analysis for RGS5, inflammatory markers (MCP-1 and VCAM-1) and macrophage-like markers (C3, CD74 and Lyz2) in mouse VSMCs transfected with si-Con (50 nM) or si-RGS5 (50 nM) followed TNF-α treatment for 48 h. (B) Representative immunofluorescence images of CD74 (green) stained in mouse VSMCs transfected with siRNA (si-Con) (50 nM) or siRNA-RGS5 (si-RGS5) (50 nM) followed TNF-α treatment for 72 h. Nucleuses stained by DAPI were indicated in blue. Scale bar = 75 μm. (C) Phagocytosis of neutral red by VSMCs transfected with si-Con (50 nM) or si-RGS5 (50 nM) followed TNF-α treatment for 7 days. (D) Phagocytosis of fluorescent microsphere by VSMCs transfected with si-Con (50 nM) or si-RGS5 (50 nM) followed TNF-α treatment for 7 days. (E, G) Representative immunofluorescence images of CD74 and α-SMA in sham-operated and ligated carotid arteries at day 14 post surgery of 10-week-old male WT mice intravenous infected with AAV-shRGS5 or 10-week-old male Rgs5-KO mice and WT littermates. Scale bar = 75 µm. (F) Western blot analysis and quantification of protein expression of RGS5 and macrophage-like markers (C3, CD74 and Lyz2) in lysates of carotid arteries harvested from 10-week-old male WT mice intravenous infected with AAV-shRGS5 following by carotid artery ligation for 14 days. (H, I) Representative cross sections of immunofluorescence or hematoxylin & eosin-staining (H&E) images of C3 and α-SMA in sham-operated and ligated carotid arteries at day 14 post surgery of 10-week-old male Rgs5-KO mice and WT littermates. Scale bar = 75 µm. The data were presented as the mean ± SD