Figure 1: MDAs enhance VSVΔ51 spread in cancer but not normal cells.

(a,b) 786-0 cancer (a) and GM-38 normal (b) cells were pretreated with MDAs (x axis) and then challenged with VSVΔ51, MOI 0.01. Forty-eight hours later, cell viability was assessed. (c) 786-0, U251 and 4T1 cancer cells as well as GM-38 were pretreated with colchicine (x axis) and infected as above for 48 h and supernatants titred. (d,e) 786-0 and GM-38 cells were treated with 100 nM colchicine or vehicle before infection with VSVΔ51-GFP or wtVSV, as above. At 48 h post infection, phase contrast and fluorescent images were taken (e, scale bar, 50 μm), and cell lysates subject to western blot, where VSV proteins are indicated by arrows (d). (f,g) 786-0 (f) and GM-38 (g) were treated with 0 (Control) or 100 nM colchicine for 24 h and fixed and probed for β-tubulin (yellow) and nuclei stained with DAPI (blue). Objective (× 100), scale bar, 20 μm, white arrow denotes polynucleation. (h) 786-0s were treated as in d and 48 h after infection, nonpermeabilized cells were stained with PI and sorted by flow cytometry. (i) 786-0s were pretreated with colchicine as above (solid line/white squares) or vehicle (dotted line/black circles) and infected with MOI 0.01 (multistep) or MOI 3 (single step) of VSVΔ51 or wtVSV, and supernatants were titred by plaque assay. For a–c, h–i, error bars or numbers represent s.e. from at least three independent experiments.