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Desmuslin gene knockdown causes altered expression of phenotype markers and differentiation of saphenous vein smooth muscle cells  Ying Xiao, PhD, Zhibin.

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Presentation on theme: "Desmuslin gene knockdown causes altered expression of phenotype markers and differentiation of saphenous vein smooth muscle cells  Ying Xiao, PhD, Zhibin."— Presentation transcript:

1 Desmuslin gene knockdown causes altered expression of phenotype markers and differentiation of saphenous vein smooth muscle cells  Ying Xiao, PhD, Zhibin Huang, MD, Henghui Yin, PhD, Hui Zhang, PhD, Shenming Wang, PhD  Journal of Vascular Surgery  Volume 52, Issue 3, Pages (September 2010) DOI: /j.jvs Copyright © Terms and Conditions

2 Fig 1 Desmuslin downregulation by short interfering RNA (siRNA) in vascular smooth muscle cells (VSMCs). VSMCs were transfected with desmuslin siRNA oligos 1 or 2 or siRNA scramble. A subset of cells was also mock transfected. Desmuslin protein levels were assessed by Western blot and normalized with reference to α-tubulin levels. Representative blots are shown. The results are presented as mean ± standard deviation as determined from three independent experiments (ie, cells were obtained from three different patients). *P < .05 vs scramble; **P < .05 vs mock; ***P < .05 vs oligo 1. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

3 Fig 2 Expression of vascular smooth muscle cell phenotype-dependent markers (smooth muscle [SM] α-actin, smooth muscle myosin heavy chain [SM-MHC], and smoothelin) following desmuslin gene knockdown by short interfering RNA (siRNA). Control cells were transfected with siRNA scramble. Protein levels were assessed by Western blot and normalized with reference to α-tubulin levels. Representative blots are shown. The results are presented as mean ± standard deviation as determined from five independent experiments (ie, cells were obtained from five different patients). *P < .05 vs scramble. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

4 Fig 3 Expression of matrix metalloproteinase-2 (MMP-2) in vascular smooth muscle cells following desmuslin gene knockdown by short interfering RNA (siRNA). Control cells were transfected with siRNA scramble. Protein levels were assessed by Western blot and normalized with reference to α-tubulin levels. Representative blots are shown. The results are presented as mean ± standard deviation from five independent experiments (ie, cells were obtained from five different patients). *P < .05 vs scramble. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

5 Fig 4 Collagen synthesis in vascular smooth muscle cells after desmuslin gene knockdown by short interfering RNA (siRNA) as indicated by incorporation of L-[3,4-3H] proline into the cell layer (A) and media (B). Control cells were transfected with siRNA scramble. The results are presented as the median and interquartile range as determined from four independent experiments (ie, cells were obtained from four different patients). *P < .05 vs scramble. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

6 Fig 5 Representative immunofluorescence micrographs of vascular smooth muscle cells transfected with scramble small inhibitory RNA (siRNA) and desmuslin siRNA. Cells transfected with siRNA scramble exhibited parallel and dense actin filaments, while cells treated with the desmuslin siRNA had dispersed and thinner filaments. Cells transfected with siRNA scramble retained a normal spindle-like morphology, while cells transfected with desmuslin siRNA were more polygonal in shape. Desmuslin-knockdown cells were larger than control cells. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

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