Journal: Journal of Biological Chemistry
Article Title: Cloning of an Immunoglobulin Family Adhesion Molecule Selectively Expressed by Endothelial Cells
doi: 10.1074/jbc.m100630200
Figure Lengend Snippet: FIG. 2. Northern blot analysis of ESAM expression in murine tissues and cultured cells. Murine and human cDNA fragments were employed as probes for hybridization to blots containing total RNA isolated from murine adult tissues and murine and human cultured cells. A dominant band corresponding to a primary ESAM transcript was observed at 2.1 kilobases in both murine and human tissues, with size determined by comparison to the mobility of 28 S and 18 S riboso- mal RNA. A second larger band was observed in cultured cells that expressed the 2.1-kilobase ESAM transcript and may represent un- spliced RNA species, alternatively spliced mRNAs from the ESAM locus, or transcripts from different genes with significant nucleotide similarity. Blots were made with total RNAs (20 mg) isolated from murine tissue and from HUVEC, human coronary artery endothelial cells (HCAEC), Namalwa (human B-cell malignancy), Molt4 (human T-cell malignancy), JEG-3 (human choriocarcinoma cells), HeLa (hu- man epithelial cell tumor), 143B (human osteosarcoma cells), A549 (human lung epithelial cell), HepG2 (human hepatocyte) cells, MEG01 (human megakaryocytic leukemia cell line), human aortic smooth mus- cle cells (HAoSMC), human coronary artery smooth muscle cells (HCASMC), human epidermal keratinocyte (NHEK), 4MBr-5 (lung ep- ithelial cell line), hemangioendothelioma (EOMA), Py-4-1 (mouse endo- thelial cell line), MEL (mouse erythroleukemia cell line), RAW (mouse monocyte/macrophage cell line), NIH3T3 (mouse embryonic fibroblast cell line), C2C12 (mouse myoblast cell line), MH-S (mouse alveolar macrophage), WEHI (mouse monocyte/macrophage cell line), MES13 (mouse mesangial cell line), TCMK (mouse kidney epithelial cell line), Neuro2a (mouse neuroblastoma cells), mouse aortic smooth muscle cells (AoSMC). A cyclophilin (Cyph) probe was used as a control for assessing RNA loading.
Article Snippet: Cloning by Suppression Subtraction Hybridization—Suppression subtraction hybridization, a polymerase chain reaction-based cDNA subtraction method (CLONTECH, Palo Alto, CA), was employed to identify genes preferentially expressed in tube-forming endothelial cells as previously described (21).
Techniques: Northern Blot, Expressing, Cell Culture, Hybridization, Isolation, Comparison, Control
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