Additional file 5 of Cooperative treatment effectiveness of ATR and HSP90 inhibition in Ewing’s sarcoma cells

Additional file 5: Figure S5. Analysis of ER stress. WE-68 and A673 cells were treated with 15–45 nM of AUY922, 2 µM of VE821, 5 µM of KU55933 and their combinations for 24 h. (A) Intracellular reactive oxygen species (ROS) level were analyzed by flow cytometry using CM-H2DCFDA. Both graphs show the mean ± SEM of three independent experiments. (B) Quantification of LAMP1 analyzed by Western blot (see Fig. 4) is shown representing the mean ± SEM of two independent experiments. (C) Otherwise isogenic p53 wild-type (wt) and p53 null (p53-/-) HCT116 cells were treated with 45 nM AUY922 ± 2 µM VE821 for 24 h. Analysis of indicated proteins was done by Western blot. ⍺-tubulin and vinculin were used to control protein loading. Immunoblots are representative for at least two independent experiments.