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. 2016 May 16;6:25944. doi: 10.1038/srep25944

Figure 1. Lentiviral NOR-1 over-expression induces phenotypic changes in human VSMC and up-regulates SMPX.

Figure 1

VSMC were transduced with lentiviral vectors to over-express NOR-1 (pLVX/NOR-1; pNOR-1), a FLAG-tagged derivative form of NOR-1 (pLVX/NOR-1-FLAG; pNOR-1F) or with control lentiviral vectors (pLVX or the EGFP expression vector [pLVX/EGFP; pGFP]). (a) Immunofluorescence microscopy analysis showing FLAG-tagged NOR-1 (green), filamentous actin (F-actin stained with phalloidin, red) and nuclei (Hoechst stain, blue). (b) Expression levels (analyzed by real-time PCR) of a panel of genes related to cell shape/morphology encoding for cytoskeleton and cytoskeletal-associated proteins in control cells (white bars) and cells transduced with pNOR-1F (black bars): ACTA2 (smooth muscle α-2 actin), MYH10 (myosin heavy chain 10), TPM1 (tropomyosin 1), TAGLN (transgelin), CALD1(1) (caldesmon transcript specific for smooth muscle cells), CALD1(2) (caldesmon transcript ubiquitously expressed, encoding for a light form of the protein [l-CaD]), ACTB (β-actin), PXN (paxillin) and SMPX (small muscle protein, X-linked) (n = 6). *P < 0.0001 vs. cells transduced with pGFP. (c) CALD1(2) and SMPX mRNA levels in VSMC transduced with pNOR-1 or with control lentiviral vectors. Data were normalized as in (b) (n = 6). *P < 0.0001 vs. cells transduced with pLVX. (d) Representative Western blot showing protein levels of NOR-1, SMPX, l-CaD, MYH10 and β-actin in VSMC over-expressing NOR-1 and control cells (n = 4).