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Blood, Vol. 106, Issue 1, 86-94, July 1, 2005
VCAM-1 expression in adult hematopoietic and nonhematopoietic cells is controlled by tissue-inductive signals and reflects their developmental origin
Blood Ulyanova et al.
106: 86
Supplemental materials for: Ulyanova et al, Vol 106, Issue 1, 86-94
Files in this Data Supplement:
- Table S1. Proportions of lineage-affiliated cells in BM and PB from f/f and Δ/Δ mice at 14 or more than 40 weeks of age (PDF, 101 KB)
- Table S2. Cell-cycle analysis of bone marrow and spleen before and during infusion of G-CSF and Flt-3 ligand (% in S-phase) (PDF, 72.3 KB)
- Figure S1. Endothelial-cell cultures from bone (PDF, 475 KB) -
All photographs were taken with a Nikon COOLPIX 995 digital camera with an adapter that fit into the opening of the eyepiece of the microscope. (A) Morphology of adherent endothelial cells (10×/0.25 objective lens on an Olympus CK2 inverted-phase microscope [Olympus America, Melville, NY]). (B) Colonies forming tubes in Matrigel cultures following manufacturer’s instructions (BD Biosciences, Bedford, MA). Images in panel B were obtained on the same microscope as for panel A, with 4×/0.1 (left panel) and 10×/0.25 (right panel) objective lenses. (C) VectaShield-mounted diI-Ac-LDL labeling of endothelial-cell cultures (red). The right panel is additionally stained with biotinylated Isolectin GSL-1 B4 and Streptavidin 488 (green). Images were obtained with a Nikon Eclipse E800 fluorescent microscope equipped with a 20×/0.40 objective lens.
- Figure S2. Tie2 and VCAM-1 expression in bone marrow fibroblast cultures (PDF, 68.6 KB) -
(A) Total RNA was isolated from the bone marrow cells as well as from established bone marrow fibroblast cultures from VCAM-1f/f and VCAM-1Δ/Δ mice, and was reverse-transcribed using random nonamer primer. PCR was then performed using Tie2- and HPRT-specific sets of primers. (B) VCAM-1 expression on cultured fibroblasts from control mice. Dot blot of cultured BM fibroblasts doubly labeled with anti-CD45 and anti–VCAM-1 (MK-2) antibodies. Note that CD45- cells (fibroblasts) retain VCAM-1 expression at early passages in vitro. CD45+ cells are macrophages.
- Figure S3. Immunohistochemistry of spleen sections stained with anti-VCAM-1 following G-CSF plus FL mobilization of mice (PDF, 120 KB) -
Note the intensity of VCAM-1 labeling in red pulp in the spleen of a VCAM-1f/f mouse (left panel) and the presence of VCAM-1 positivity in follicular dendritic cells (arrow) in the spleen of one VCAM-1–ablated mouse (right panel). Image acquisition was performed as described for Figure 4.
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