Fig. 3

Live cell validation of MSAP-Ac-TZ14011 staining in EWS cells. a, b MDA-MB-231 X4 containing transfected CXCR4-GFP (green) and EWS cell lines (c, d) TC32 and (e, f) IARC-EW7 were imaged by live cell confocal microscopy directly after incubation with MSAP-Ac-TZ14011 (red) (T = 0) and after 3 h incubation (T = 3). At T = 0 MDA-MB-231 X4 and TC32 demonstrated a membrane staining where no staining was observed in IARC-EW7. IARC-EW7 and TC32 were half an hour prior to imaging incubated with lysotracker DND-26 (green). Hoechst staining (blue) was used to stain the nucleus. All images were taken using a 63× objective. In the MDA-MB-231 × 4 cell line at T = 3 intracellular MSAP-Ac-TZ14011 signal was detected that was co-localized with CXCR4-GFP signal (panel b and inset). In TC32 cell the lysotracker DND-26 signal was co-localized with the MSAP-Ac-TZ14011 signal (d, arrow and inset)