Skip to main content
. Author manuscript; available in PMC: 2016 Sep 1.
Published in final edited form as: J Immunol. 2015 Jul 22;195(5):2461–2471. doi: 10.4049/jimmunol.1500105

Figure 4. IFN-β Suppresses Transcriptional Responses to Purified Ligands Across Multiple Innate Receptor Systems.

Figure 4

(A) WT C57BL6/J macrophages were pretreated for 60 minutes either with media alone or 100 U/ml IFN-β. Macrophages were stimulated with 50 ng/ml purified E.coli LPS for the indicated times. RNA was harvested and transcript levels assayed by qRT PCR. (B) Macrophages were pre-treated as in (A) and stimulated with 50 ng/ml LPS for 6 hours. Whole cell lysates were probed by western blot for pro-IL1β protein. (C) Samples of cell supernatants from macrophages treated as in (A) were collected at a 24 hour time-point and assayed for chemokine content by ELISA. (D) WT C57BL6/J peritoneal macrophages were pretreated with 100 U/ml mammalian IFN-β for the indicated times and subsequently re-stimulated with 50 ng/ml E. coli LPS for 6 hours. RNA was harvested and IL-1β transcript expression quantified by qRT PCR. (E) WT C57BL6/J peritoneal macrophages were pretreated with 100 U/ml mammalian IFN-β for 60 minutes prior to stimulation for the indicated times with the TLR2 ligand Pam3Cys (250ng/ml). (F) Peritoneal macrophages pre-treated with media alone or IFN-β were subsequently treated overnight with the NOD-1 ligand C12-iE-DAP (20μg/ml) or the NOD-2 ligand L18-MDP (20μg/ml). Samples of cell supernatant were harvested and CXCL1 levels assayed by ELISA. Data in A-F is representative of three independent experiments. Data is shown as mean ± SEM.