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. 2019 Mar 20;124(9):1309–1322. doi: 10.1161/CIRCRESAHA.118.314187

Figure 6.

Figure 6.

SLC1A5 (solute carrier family 1 member 5) activates mTORC1 (mechanistic target of rapamycin complex 1) signaling and promotes vascular smooth muscle cell proliferation. A, Human coronary artery smooth muscle cells (HCASMCs) were transduced with GFP (green fluorescent protein) or SLC1A5 adenovirus for WST-1 (water soluble tetrazolium-1) proliferation assay at day 3 post-viral transduction. B, Immunofluorescence for EdU (5-ethynyl-2′-deoxyuridine; red) was performed to assess HCASMC proliferation after GFP or SLC1A5 viral transduction for 3 d, and (C) the percentages of EdU+ cells to total cell numbers were quantified. Cell nuclei were counterstained with DAPI (4’,6-diamidino-2-phenylindole; blue). D, Numbers of HCASMCs were counted post 3 d of GFP or SLC1A5 viral transduction. n=5. E, Western blot was performed to examine the effects of myc-tagged SLC1A5 overexpression on mTORC1 signaling activation and smooth muscle contractile gene expression in HCASMCs. F, HCASMCs were transfected with scrambled control siRNA (siControl) or SLC1A5 siRNA (siSLC1A5) for WST-1 proliferation assay or (G) cell count analysis at day 3 post-transfection. n=3 per group. H, HCASMCs were transfected with control siRNA (siControl) or SLC1A5 siRNA (siSLC1A5) for 3 d and then cell lysates were harvested for Western blot analysis. L indicates long exposure; and S, short exposure. *P<0.05.