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1.
Figure 2.

Figure 2. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

FACS analysis of chemokine receptor transfectants with anti-D6 antibodies. Approximately 10 6 L1.2 cells stably expressing human chemokine receptors CCR1–5 or D6 were incubated with anti-D6 antibodies (50 μl of hybridoma supernatant from clone 4A5), and then antibody-containing complexes detected using a fluorescein isothiocyanate-coupled goat anti-mouse IgG and FACS analysis.

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
2.
Figure 1.

Figure 1. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

In situ binding of radioiodinated MIP-1αP to lymphatic vessels in normal human skin. Cubes of viable human skin were incubated with 125I-MIP-1αP, sectioned, and the bound protein visualized as outlined in Materials and Methods. Tissue-bound 125I-MIP-1αP appears as black autoradiographic grains. Sections were counterstained with hemalaun. Thin arrows mark vessels with clear morphological features of lymphatics. Thick arrow indicates a resident dermal cell also labeled with 125I-MIP-1αP. The asterisk marks an unlabeled blood vessel. Original magnification, ×1,400.

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
3.
Figure 6.

Figure 6. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

D6 immunoreactivity in secondary lymphoid organs. Representative images of paraffin-embedded sections from secondary lymphoid organs, immunostained with either anti-D6 (A–D and F), or anti-podoplanin (E and G), antibodies. Immunoreactivity is visualized using an avidin-biotin-peroxidase detection system and a peroxidase substrate producing a red stain; all sections counterstained with hematoxylin. A and B: D6 immunostaining in tonsils. Asterisk marks an HEV (both original magnifications, ×1,000). C: D6-immunoreactive vessel in red pulp of a spleen (original magnification, ×1,250). D–G: Lymph node. Adjacent sections showing subcapsular (D and E) and medullary (F and G) sinuses of the lymph node stained with anti-D6 (D and F) or anti-podoplanin (E and G) antibodies. Original magnifications: ×720 (D–G).

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
4.
Figure 4.

Figure 4. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

D6 mRNA expression by lymphatic ECs in the skin. Images taken from sections of human skin hybridized to DIG-labeled sense (A), or antisense (B, C, and E), D6 RNA and visualized with alkaline phosphatase-conjugated anti-DIG antibodies and BCIP/NBT solution. The presence of hybridized probe is indicated by a blue stain. D and F are sections adjacent to those used in C and E, respectively, and are immunostained with anti-podoplanin antibodies, visualized with an alkaline phosphatase-anti-alkaline phosphatase kit producing a red stain, and counterstained with hemalaun. A–D: Images taken from the upper dermis, with epidermis visible. B (inset), E and F: Images of deep dermal lymphatic vessels. Original magnifications: ×240 (A–D); ×100) (E and F).

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
5.
Figure 7.

Figure 7. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

D6 immunoreactivity in vascular tumors. Paraffin-embedded sections of vascular tumors stained with anti-D6 antibodies (A–E), or anti-podoplanin antibody (F) with immunoreactivity visualized using an avidin-biotin-peroxidase detection system and a peroxidase substrate producing a red stain. All sections counterstained with hematoxylin. A, B, C, D, and E show tumors listed in Table 1 ▶ as numbers 1, 2, 3, 6, and 11, respectively. F shows a section adjacent to E stained with anti-podoplanin antibody. Only a small proportion of podoplanin-positive cells is D6-positive. Original magnifications: ×140 (A and C); ×300 (B); ×400 (D, E, and F).

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
6.
Figure 3.

Figure 3. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

D6 immunoreactivity on lymphatic ECs in skin. Frozen sections of human skin immunostained with anti-D6 antibodies (A and D) or anti-podoplanin antibodies (C). In B, D6 immunoreactivity was detected on paraffin sections. Antibodies are visualized using an alkaline phosphatase-anti-alkaline phosphatase kit producing a red stain; all of the sections are counterstained with hemalaun. A: ECs lining two lymphatic capillaries in upper dermis display D6 immunoreactivity. Original magnification, ×2,000. B: Large collective lymphatic is stained by anti-D6 antibody, “a” marks an artery, “v” a vein, both immunonegative. Original magnification, ×900. Two insets contain resident non-ECs that show D6 immunoreactivity (both original magnifications, ×1,400). The asterisk indicates the epidermis. C and D are taken from adjacent serial sections and show a lymphatic vessel with podoplanin and D6 immunoreactivity, respectively. Original magnifications, ×1,200.

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.
7.
Figure 5.

Figure 5. From: The ?-Chemokine Receptor D6 Is Expressed by Lymphatic Endothelium and a Subset of Vascular Tumors.

D6-immunoreactive lymphatic vessels in the gut. Paraffin-embedded sections of the gut immunostained with anti-D6 antibodies visualized using an avidin-biotin-peroxidase detection system and a peroxidase substrate producing a red stain. Representative D6 immunoreactivity associated with lymphatics in a longitudinal section of a small intestinal villus (A), lamina propria mucosae of the colon (B), a cross-section of two villi in the large intestine (C), muscular layer of the colon (D), lymphoid follicle in the lamina propria (E), lamina muscularis in normal appendix (F), and a patient with acute appendicitis (G). Original magnifications: ×400 (A), ×280 (B), ×530 (C), ×600 (D), ×440 (E, F, and G). The asterisk in D marks an immunonegative blood vessel. All sections counterstained with hematoxylin.

Robert J. B. Nibbs, et al. Am J Pathol. 2001 March;158(3):867-877.

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