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. Author manuscript; available in PMC: 2021 Feb 1.
Published in final edited form as: J Leukoc Biol. 2020 May 22;109(2):309–325. doi: 10.1002/JLB.4HI0420-285RR

Figure 4. CD40 signaling through TRAF6 is required for CD40 restriction.

Figure 4.

A-B) Pmacs were harvested from male Ifnar−/− mice and stimulated with agonistic CD40 antibody in the presence of varying concentrations of a TRAF6 inhibitor (7651589) for 7 hours. Following stimulation, RNA was isolated and IL-12p35 mRNA was quantified by qRT-PCR (A). In parallel, cells were stimulated for 24 hours with CD40 agonist (+/− 10 μM TRAF6 inhibitor 7651589) and subsequently challenged with rVSV/EBOV GP (MOI=1). RNA was harvested and viral RNA was assessed by qRT-PCR (B). C) Peritoneal cells were harvested from male Ifnar−/− mice and transfected with 3 TRAF6 siRNAs (25 nM each). At 48 hours, cells were stimulated with agonistic CD40 antibody or IL-12. After 24 hours cells were washed to remove non-adherent cells, media was refreshed, and cells were infected with rVSV/EBOV GP (MOI=1). RNA was harvested 24hpi and virus was quantified by qRT-PCR.