Figure 5.

Sulforaphane Dampens the Innate Antiviral Response

(A) HEK293T cells were pretreated for 24 hr with increasing doses of SFN (1–20 μM), and the ISRE promoter activity was assessed using a luciferase assay in the presence or absence of VSVΔ51 (MOI 0.01). (B) PC-3 cells were pretreated with sulforaphane (SFN) (20 μM) for 24 hr and were subsequently infected with VSVΔ51-GFP (MOI 1). Antiviral and inflammatory gene expression levels were assessed by high-throughput qPCR. (C) PC-3 cells were pretreated for 24 hr with SFN (20 μM) and subsequently challenged with VSVΔ51-GFP (MOI 1) for 24 hr. Whole-cell extracts were analyzed for P-IRF3, IRF3, P-STAT1, STAT1, STING, HO-1, LC3, VSV proteins, and β-actin by immunoblotting. (D) PC-3 cells were pretreated for 24 hr with SFN (20 μM) and subsequently challenged with VSVΔ51-GFP (MOI 1) for 8 hr. Translocation of IRF3 and Nrf2 from cytoplasm to nucleus was investigated by immunoblotting in cytoplasmic and nuclear fractions at the same time. The experiment was repeated twice.