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Volume 144, Issue 1, Pages 7-14 (May 1999)

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1 Volume 144, Issue 1, Pages 7-14 (May 1999)
Concordant upregulation of type II-TGF-β-receptor, the cyclin-dependent kinases inhibitor P27Kip1 and cyclin E in human atherosclerotic tissue: implications for lesion cellularity  Christian Ihling, Katja Technau, Volkmar Gross, Jürgen Schulte-Mönting, Andreas M. Zeiher, Hans E. Schaefer  Atherosclerosis  Volume 144, Issue 1, Pages 7-14 (May 1999) DOI: /S (99)

2 Fig. 1 Double-immunostaining for p27 (anti-p27 monoclonal or polyclonal, blue reaction-product, APAAP method, CD3 (anti-CD3, polyclonal, brown reaction-product, ABC method, panel A), CD68 (anti CD68 monoclonal, brown reaction product, ABC method, panel B), and α-actin, monoclonal, brown reaction-product, panels C and D) showing numerous T-lymphocytes, macrophages, vascular smooth muscle cells and microvascular endothelial cells with nuclear expression of p27. Magnification: A, B, C and D ×350. Atherosclerosis  , 7-14DOI: ( /S (99) )

3 Fig. 2 Semi-serial sections of a human carotid endarterectomy specimen with advanced complicated atherosclerosis showing a plaque area with numerous vascular smooth muscle cells. Low-power magnification of (A) p27 (anti-p27, monoclonal, dark-brown reaction-product) and TGF-β-RI (anti- TGF-β-RI, polyclonal, red reaction-product) double immunolabeling, and (B) p27 (anti-p27, monoclonal, red-brown reaction-product) and TGF-β-RII (anti- TGF-β-RII, polyclonal, blue reaction- product) double immunolabeling with positive nuclear staining for p27 and positive membranous and cytoplasmic staining for TGF-β-RI and TGF-β-RII, respectively. Note that nearly all cells demonstrate coexpression of p27, TGF-β-RI and TGF-β-RII. The insets in A and B show higher powers from panels A and B, respectively, demonstrating TGF-β-RI (panel A, red reaction-product) as well as TGF-β-RII (panel B, blue reaction-product) in a membraneous and a diffuse cytoplasmic form. Magnification: A and B ×200, inset A and B ×1000. Atherosclerosis  , 7-14DOI: ( /S (99) )

4 Fig. 3 Semi-serial sections of a human coronary artery with normal histology. Low-power magnification of (A) p27 (anti-p27, monoclonal, black reaction-product) and TGF-β-RI (anti TGF-β-RI, polyclonal, red reaction product) double immunolabeling, and (B) p27 (anti-p27, monoclonal, brown reaction product) and TGF-β-RII (anti- TGF-β-RII, polyclonal, blue reaction-product) double immunolabeling with positive nuclear staining for p27 and positive membranous and cytoplasmic staining for TGF-β-RI and TGF-β-RII, respectively. Note in panel A that most of the medial SMCs show coexpression of p27 and TGF-β-RI whereas TGF-β-RII staining on panel B is less numerous. The insets in A and B show higher powers from panels A and B, respectively, demonstrating TGF-β-RI (panel A, red reaction-product) as well as TGF-β-RII (panel B, blue reaction-product) in a membraneous and a diffuse cytoplasmic form. Note on the inset in panel B that there are several cells without TGF-β-RII staining. Magnification: A and B ×200, inset A and B ×1000. Atherosclerosis  , 7-14DOI: ( /S (99) )

5 Fig. 4 Double-immunostaining for cyclin E (brown reaction-product, ABC method, panel A) and CD68 (blue reaction-product, APAAP method) showing numerous macrophages with nuclear staining for CE and cytoplasmic staining for CD68. Double-immunostaining for CE (brown reaction-product, ABC method, panel B) and α-actin (blue reaction-product, APAAP method) showing intimal vascular smooth muscle cells with nuclear staining for CE and cytoplasmic staining for α-actin. Magnification: A and B ×350. Atherosclerosis  , 7-14DOI: ( /S (99) )

6 Fig. 5 Double immunofluorescence labeling for P27 (anti-P27 polyclonal, green fluorescence) and CE (anti-CE, monoclonal red fluorescence). Nuclei were counterstained with DAPI (blue fluorescence). There are few intimal cells showing nuclear immunostaining for p27 and cyclin E. However, most of the cells show positive nuclear staining for p27 (green fluorescence) in the absence of CE staining. Panel A indicates red fluorescence of CE staining. Note that there is a weak and finely granular red fluorescence in the nuclei (compare with panel C) on a background of diffuse red fluorescence which is caused by the non-specific autofluorescence of the lipopigment present in the atherosclerotic tissue. Panel B indicates green fluorescence of p27 staining. Panel C indicates blue fluorescence of nuclear DAPI staining. Double-labeled cells are marked by arrows. Magnification: A, B and C ×875. Atherosclerosis  , 7-14DOI: ( /S (99) )


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