Figure 4.

Application of dual stress drives Ewing sarcoma matrix degradation capabilities. Representative images of FITC gelatin matrix degradation assays in (A) CHLA10 cells when exposed to no stress (top panels), single stresses (middle panels) or combined serum deprivation (SFM) and hypoxia (1% O₂, bottom panels) for 24 hours. Corresponding cell staining for cortactin (red) is included as a marker for punctate invadopodia structures. Black “holes” in the green gelatin represent areas of degradation. (B) Representative images of gelatin degradation in A673 and CHLA25 cells in unstressed (FBS, 21% O₂) versus dual stress (SFM and 1% O₂) conditions are shown in the top two panel rows. Bottom panels are magnified images of the dual stress condition with the cortactin overlay included to demonstrate the overlap of punctate structures with areas of matrix degradation. Assays were performed at least in triplicate.