Type III IFN mediated expression kinetics of ISGs are independent of differential levels of IFNLR1 receptor.

Wild-type T84 cells were transduced with rIFNLR1-GFP to create a stable line overexpressing IFNLR1 tagged with GFP. (A) WT cells overexpressing IFNLR1-GFP (WT+IFNLR1-GFP) from the same population were separated by cell sorting into three populations: non (neg)-, low- and high-expressing GFP cells. Gates were created based on the auto-fluorescence of WT cells. (B) WT and WT+IFNLR1-GFP cells were treated with type III IFN (λ1−3) (300 ng/mL equivalent 13.7 nM) for 3, 6, 12 and 24 hours prior to sorting in neg-, low- and high-expressing IFNLR1-GFP cells. The kinetic pattern of expression of one representative ISG from each temporal expression patterns groups 1–4 was analyzed by qRT-PCR in each sorted population. Data are normalized to HPRT1 and are expressed relative to untreated cells at each time point. A representative experiment with technical triplicates, out of two independent experiments is shown. Mean values and SD are shown.