Figure 3
Downstream markers of EWSR1-FLI1 activity confirm the presence of EWSR1-FLI1low subpopulation. (a) Immunofluorescence of LOX (EWSR1-FLI1-downregulated target) and actin cytoskeleton in A673 and SK-N-MC cell lines. Images were acquired using upright widefield Apotome microscope (Zeiss, Marly-Le-Roi, France) equipped with a Coolsnap HQ2 camera through a x63 NA 1.4 oil-immersion objective lens and driven by Axiovision software (Zeiss). White arrows point the cells of interest, exhibiting LOX expression associated to an important cell-spreading and a well-organized actin stress fibre cytoskeleton. Scale bars represent 20 μm. (b) IHC of LOX in shA673-1c+/−DOX xenografts. Black arrows indicate the cells of interest. Right panels represent enlargement of left panels. Scale bars represent 50 μm. (c) IHC and quantification of LOX in eight representative samples from Ewing tumours TMA. Black arrows point cells expressing LOX. Scale bars represent 20 μm. Error bars mean s.d.
