Talin and vinculin are downregulated in atherosclerotic plaque; Tampere Vascular Study Magdaléna von Essen, Rolle Rahikainen, Niku Oksala, Emma Raitoharju,

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Talin and vinculin are downregulated in atherosclerotic plaque; Tampere Vascular Study Magdaléna von Essen, Rolle Rahikainen, Niku Oksala, Emma Raitoharju, Ilkka Seppälä, Ari Mennander, Thanos Sioris, Ivana Kholová, Norman Klopp, Thomas Illig, Pekka J. Karhunen, Mika Kähönen, Terho Lehtimäki, Vesa P. Hytönen Atherosclerosis Volume 255, Pages 43-53 (December 2016) DOI: 10.1016/j.atherosclerosis.2016.10.031 Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Expression of talin (TLN) and vinculin (VCL) transcripts. Results of microarray analysis (MA). (A–D) Expression levels in LITA controls and all, stable and unstable atherosclerotic plaques. (E–H) Expression levels in carotid, abdominal aortic and femoral arterial bed compared to LITA controls. Atherosclerosis 2016 255, 43-53DOI: (10.1016/j.atherosclerosis.2016.10.031) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Heat maps of log2 expression values. Talin-1 (TLN1), talin-2 (TLN2) and vinculin (VCL) co-expression with biomarkers of inflammation (CD68, ALOX5) and smooth muscle cells markers (CNN1, SMTN). The expression values of each row (gene) are scaled to z-scores, color-code for expression values and arterial site is presented in the top-right corner. The dendrogram illustrates hierarchical clustering based on the seven robustly expressed genes. The top bar represents the arterial site origin of the sample tissue. Atherosclerosis 2016 255, 43-53DOI: (10.1016/j.atherosclerosis.2016.10.031) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Tissue localization of talin-1, talin-2 and vinculin in atherosclerotic plaque from carotid artery and left internal thoracic (LITA) control. (A) Talin-1 and PECAM-1, (B) talin-1 and talin-2, (C) vinculin and PECAM-1. Talin-1 was expressed in the endothelial cells of LITA, while talin-1 expression in plaque endothelial cells was not observed (A). Talin-2 expression was decreased in plaque endothelium as compared to LITA, but not as strongly as talin-1 expression (B). In LITA, high vinculin expression was observed in tunica intima and at the lumenal side of tunica media. In plaque samples, decreased vinculin expression was observed in the thickened tunica intima (C). PECAM-1 expression was found to be increased in plaque endothelium (A, C). L, lumen; I, tunica intima; E1, internal elastic lamina; E2, external elastic lamina; M, tunica media; A, tunica adventitia. Atherosclerosis 2016 255, 43-53DOI: (10.1016/j.atherosclerosis.2016.10.031) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Tissue morphology and localization of talin-1 in immunostained FFPE sections of LITA from patients with coronary artery disease and in sections of healthy abdominal aorta and carotid artery. Uniform talin-1 immunostaining was observed in tunica intima and tunica media in both immunostained LITA samples, as well as in the samples from healthy abdominal aorta and carotid artery. HE-staining showed normal tissue morphology for all of the analyzed samples. In the merged images, talin-1 immunostaining is shown as green and DAPI chromatin staining as blue. L, lumen; I, tunica intima; E1, internal elastic lamina; E2, external elastic lamina; M, tunica media; A, tunica adventitia. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Atherosclerosis 2016 255, 43-53DOI: (10.1016/j.atherosclerosis.2016.10.031) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Simplified speculative model for downregulation of talin expression in vascular endothelium as initial trigger of atherosclerotic plaque formation. Atherosclerosis 2016 255, 43-53DOI: (10.1016/j.atherosclerosis.2016.10.031) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Atherosclerosis 2016 255, 43-53DOI: (10. 1016/j. atherosclerosis. 2016 Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions