Fig 2: EndoC-βH1 were treated with IFNα (1000 U/mL) or IFNγ (20 ng/mL) or IFNʎ1 or 2 (200 ng/mL) for 24 h. After this period, cells were lysed and 2% of the input lysate removed, denatured and stored at −20°C. The remainder of the protein lysate was incubated with 2 µg of STAT1 antibody or 2 µg of isotype control IgG overnight at 4°C. A 50% bead slurry was added, proteins eluted and Western blotting was performed using antisera directed against either STAT2 and STAT1.

Fig 3: EndoC-βH1 cells were treated with various interferons (IFNα – 1000 U/mL; IFNɤ – 20 ng/mL; IFNʎ1 or 2 – 200 ng/mL) for either 30 min or 24 h. Protein was extracted at the end of the incubation and Western blotting performed using anti-phospho STAT1, anti STAT1, anti-phospho STAT2 and anti STAT2. A representative blot is shown for each target protein, but densitometric traces were obtained from a minimum of three separate experiments in each case (lower panels). GAPDH was used as a loading control. ****P < 0.0001; **P < 0.01; *P < 0.05.

Fig 4: EndoC-βH1 treated with either 1000 U/mL IFNα, 20 ng/mL IFNγ or 200 ng/mL IFNʎ1 or IFNʎ2 for 30 min. Cells were fixed with 4% paraformaldehyde, permeabilised and stained with anti-STAT1 or anti-STAT2 antisera, as shown. Images were taken using a Leica DM4000 B LED Fluorescence microscope using exposure settings defined for the control conditions (upper and middle panels of each figure). In the lower panels of each figure, the exposure settings were adjusted to provide improved resolution of the subcellular localisation of each STAT isoform after incubation with interferons.

Fig 5: EndoC-βH1 were treated with either IFNα (1000 U/mL) or IFNλ1 or IFNλ2 (20 ng/mL) for 24 h. After this time, cells were washed and retreated with either IFNα (1000 U/mL) or IFNλ1 or IFNλ2 (20 ng/mL) for a further 30 min. Protein was extracted at the end of the incubation and Western blotting performed using anti-pSTAT1, anti-total STAT1, anti-pSTAT2 and anti-total STAT2. GAPDH was used as a loading control (representative blots are shown; n = 3).