Journal:
Article Title: Receptor Density Is Key to the Alpha2/Beta Interferon Differential Activities ▿
doi: 10.1128/MCB.01808-08
Figure Lengend Snippet: (A) Graphic representation of the relative antiproliferative potencies of IFN-α2 and IFN-β in the human cell lines 2fTGH (fibrosarcoma), WISH (amniotic), MDA231 (breast cancer), and Daudi (B lymphoma) and the three U5-derived clones (U5-low/low, U5-low/hi, and U5-hi/hi) described in this study. EC50s (pM) for IFN-α2 are plotted against the ratio of the EC50 of IFN-α2 to that of IFN-β for each cell line (Table (Table1).1). (B) Levels of IFNAR1 and IFNAR2 in the three U5-derived clones (U5-low/low, U5-low/hi, and U5-hi/hi) and in the parental 2fTGH cells. Surface IFNAR1 and IFNAR2 were quantified by FACS analysis using MAbs AA3 and CD118, respectively. Dark gray area, isotypic control; light gray area, 2fTGH; black line, U5-low/low, U5-low/hi, or U5-hi/hi. Of note, these clones exhibited similar forward scatter values, as determined by FACS analysis, indicating that the increases observed are due to changes in surface receptor density and not cell volume. (C) Analysis of the expression level of Jak/Stat pathway components in 2fTGH cells and the U5-derived clones. Total cell lysates (30 μg) were analyzed by Western blotting with the indicated Abs. Loading was evaluated by measuring Akt levels.
Article Snippet: Surface receptor levels were monitored as described in reference 18 , using monoclonal antibodies (MABs) AA3 (BiogenIdec, Boston) and CD118 (PBL, Piscataway, NJ), which are specific for IFNAR1 and IFNAR2 respectively.
Techniques: Derivative Assay, Clone Assay, Expressing, Western Blot