1. Fig. 6 DMF inhibits NF-κB translocation upon infection.
DMF inhibits NF-κB translocation upon infection. (A) Structures of DMS, MMS, and S. (B and C) cells were pretreated with the indicated DMF analogs for 4 hours and subsequently infected with oncolytic VSVΔ51 expressing GFP at an MOI of (B) At 24 hours after infection, fluorescence images were taken from the infected cells. (C) Corresponding viral titers were determined from supernatants 48 hours after infection (n = 3; mean ± SD; ***P < 0.001, one-way ANOVA, as compared to the untreated counterpart). (D) GSH concentrations were determined in cells after a 4-hour treatment with FMAEs (n = 4; mean ± SD; ***P < 0.001, one-way ANOVA, as compared to the untreated counterpart). (E) cells were grown in the presence or absence of BSO (2 mM) for 7 days and pretreated with DMF (200 μM) for 4 hours or left untreated and then infected with oncolytic VSVΔ51 expressing GFP at an MOI of Corresponding viral titers were determined from supernatants 48 hours after infection (n = 3; mean ± SD). (F) Heat map showing the expression of the differentially expressed oxidative stress genes. Expression of genes was normalized to values obtained for untreated, uninfected control. (G) HMOX1 expression quantified by quantitative polymerase chain reaction (qPCR) from cells after a 6-hour treatment with FMAEs (n = 3; mean ± SD). (H) siNRF2 knockdown cells were treated with DMF or untreated and infected as in (E). Corresponding viral titers were determined from supernatants 24 hours after infection. RNA was extracted, and the expression of NRF2 and IFITM1 genes was quantified by qPCR (n = 3; mean ± SD). (I and J) Cytoplasmic and nuclear protein fractions were extracted from cells treated with DMF (200 μM) for 4 hours and (I) subsequently infected with oncolytic VSVΔ51 expressing GFP at an MOI of 1 for 8 hours or (J) treated with TNFα (30 ng/ml) for 30 min. Cell lysates were probed for multiple proteins, as indicated, by Western blot. (K) cells were pretreated with NF-κB inhibitors targeting IKK [IKK16 (10 μM) and TPCA1 (40 μM)] for 4 hours and subsequently cotreated with DMF (150 μM) and oncolytic VSVΔ51 expressing GFP at an MOI of Corresponding viral titers were determined from supernatants 24 hours after infection (n = 3; mean ± SD; ***P < 0.001, one-way ANOVA, as compared to the untreated counterpart).
Mohammed Selman et al., Sci Transl Med 2018;10:eaao1613
Published by AAAS