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. 2017 May 17;25(8):1900–1916. doi: 10.1016/j.ymthe.2017.04.022

Figure 1.

Figure 1

Sulforaphane Enhances VSVΔ51-Mediated Oncolytic Activity in Resistant PC-3 Cells

PC-3 cells were pretreated for 24 hr with increasing concentrations of sulforaphane (SFN) and were subsequently infected with VSVΔ51-GFP (MOI 1). (A and B) Infectivity was determined by fluorescent microscopy (A) or quantified by flow cytometry (B) at the indicated times. (C) Viral replication was assessed by plaque assay. (D) Cell survival was imaged by light microscopy 72 hr following VSVΔ51 challenge. (E and F) The percentage of viable (E) and apoptotic cells (F) was assessed using an annexin-V/7AAD staining by flow cytometry at the indicated times. (A–F) Data are the means ± SEM from two experiments performed in triplicate. (G) The correlation between the percentage of VSVΔ51-GFP+ cells at 24 hr and the percentage of apoptotic annexin-V+ cells at 48 hr was calculated in PC-3 cells using a Spearman test (n = 33). (H) The same statistical test was used to calculate the correlation between the percentage of apoptotic annexin-V+ cells and the percentage of MitoSOX+ cells at 48 hr after infection (n = 36). (I) PC-3 cells were pretreated for 24 hr with SFN (20 μM) and challenged with VSVΔ51-GFP (MOI 1) for 48 hr. Whole-cell extracts (WCEs) were analyzed by immunoblotting for cleaved caspase-3, cleaved caspase-7 (high and low exposure), cleaved PARP, VSVΔ51-GFP, and β-actin. Results are from a representative experiment; all immunoblots are from the same samples.