1. Fig. 1 DMF promotes viral infection.
DMF promotes viral infection. (A) Structure of DMF. (B) VSVΔ51-resistant human renal cancer cell line was pretreated with DMF or left untreated for 4 hours and subsequently infected with VSVΔ51 (MOI, 0.01). Corresponding viral titers were determined 48 hours after infection from supernatants [n = 3; mean ± SD; *P < 0.05, ***P < 0.001, one-way analysis of variance (ANOVA), as compared to the untreated condition]. PFU, plaque-forming units. (C and D) Various human and murine cell lines were pretreated with DMF (150 to 250 μM) or left untreated for 4 hours and subsequently infected with VSVΔ51 (MOI, 0.01). Twenty-four hours after infection, fluorescence images were taken from the infected cancer cells, as shown in (C). Corresponding viral titers were determined 48 hours after infection from supernatants, as shown in (D) (n = 3 to 4; mean ± SD; P < 0.05, two-tailed t test, as compared to the untreated counterpart for each cell line). (E and F) cells were pretreated with DMF (150 μM) or left untreated for 4 hours and subsequently infected with (E) Sindbis virus (MOI, 10) or (F) HSV-1 (MOI, 0.01). Corresponding viral titers in supernatants were determined 48 hours after infection (n = 3; mean ± SD; **P < 0.01, ***P < 0.001, two-tailed t test, as compared to the untreated counterpart). (G) Human A549 cells were pretreated as in (B) and infected with an adenovirus expressing firefly luciferase (Ad5) at an MOI of 1. Luciferase activity was measured over the course of 7 days. Results are represented as relative light units (RLU), and background is indicated by a black line (n = 3; mean ± SD; P < 0.05, two-way ANOVA from day 2 to day 5). (H) Multistep (MOI, and 0.01) and single-step (MOI, 3) growth curves cells were pretreated with DMF and infected with VSVΔ51 at an MOI of 0.001, 0.01, or 3; supernatants were titered by plaque assay (n = 3; mean ± SD). (I) cells were pretreated with DMF for 4 hours and infected with VSVΔ51 (MOI, ). An agarose overlay was added after 1 hour of infection. Fluorescence microscopy of a representative plaque 48 hours after infection is shown. The complete dose range is presented in fig. S1B. Corresponding images of Coomassie blue stain of the full wells and the graph of average plaque diameters illustrate the enhancement of the plaque diameters in the presence of DMF (n = 20; horizontal lines indicate means; *P < 0.05, ***P < 0.001, one-way ANOVA, as compared to the mock-treated counterpart). (J) 786-0, CT26WT, and B16F10 cell lines were pretreated and infected as in (B). Cell viability was assayed 48 hours after infection. Results were normalized to the average of the values obtained for the corresponding uninfected, untreated cells (n = 8; mean ± SD; ***P < 0.001, one-way ANOVA, as compared to VSVΔ51 condition).
Mohammed Selman et al., Sci Transl Med 2018;10:eaao1613
Published by AAAS