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. 2011 Jul 14;6(7):e22200. doi: 10.1371/journal.pone.0022200

Figure 4. Analysis of the type I IFN receptor in naïve and primed HLLR1-1.4 cells.

Figure 4

(A) Surface level of IFNAR1 and IFNAR2 in naïve cells and in IFN β or IFN λ1-primed cells as determined by FACS. Cells were primed for 24 hr, washed and maintained in medium without IFN for 24 hr. Cells were then stained with AA3 mAb (IFNAR1) or CD118 mAb (IFNAR2) followed by biotinylated rat anti-mouse Ab and streptavidin-PE. (B) Level of phosphorylation of Stat1 in naïve and primed cells stimulated for 30 min with 100 pM of IFN α2, IFN β or IFN α2-HEQ. Lysates (30 µg) were immunoblotted with the indicated antibodies. Priming was for 8 hr followed by 16 hr resting in medium without IFN. (C and D) Binding of 125I labelled IFN α2 (C) or IFN α2-HEQ (D) at 37°C for 1 hr to naïve (closed circles), IFN β-primed cells (triangles) or IFN λ1-primed cells (open circles). Cells were primed for 8 hr and maintained without IFN for 16 hr.