Figure EV2. Quantitative data with different prestimulation doses for model calibration.

1. Model calibration with IFNα‐induced expression of feedback transcripts STAT1, STAT2, IRF9, IRF2, USP18, SOCS1, and SOCS3. Growth factor‐depleted Huh7.5 cells were prestimulated with 2.8, 28, and 1,400 pM IFNα (yellow background). After 24 h, cells were stimulated with 1,400 pM IFNα or were left untreated (white background). IFNα‐induced expression of target genes was measured by qRT‐PCR. RNA levels were normalized to the geometric mean of reference genes GAPDH, HPRT, and TBP and were displayed as fold change, visualized by filled circles with errors representing 1σ confidence intervals estimated from biological replicates (N = 3 to N = 14) using a combined scaling and error model. Model trajectories are represented by lines.
2. Growth factor‐depleted Huh7.5 were prestimulated with a range of 0.28–1,400 pM IFNα for 24 h and stimulated with 1,400 or 2,800 pM IFNα. IFNα‐induced phosphorylation of nuclear STAT1 and STAT2 and abundance of feedback proteins were analyzed by quantitative immunoblotting. Data were normalized to reference proteins Calnexin or HDAC1 and represented by filled circles with errors representing 1σ confidence intervals estimated from biological replicates (N = 1 to N = 19) using a combined scaling and error model. Model trajectories are represented by lines.