Figure 6
ONC201 pre-treatment enhances radiation response in other cancer types. (a) Western Blot analysis showing the overexpression of UPR related proteins in colorectal cancer cells HT-29 treated with ONC201 (5 and 10 µM) for 24 h, if compared to those treated with the Vehicle (representative of n = 3). The corresponding densitometric analysis are in Supplementary Fig. 11b. Here the average is expressed as a number embedded within the figure (the SD are within the 5% acceptance of statistical significance). The membranes were cut before the incubation with the primary antibody (see “Materials and methods” for specifications) and either detected alone or together. All the membranes were exposed for 3 min, with the exception of XBP1 (exposed for 7 min) and Tubulin (exposed for 2 min). (b) Cell counts of HT-29 colorectal cells primed to radiation (Xrad, 2- 8 Gy) with ONC201 for 24 h showed ONC201 impacting cells survival at 72 h. Radiation Enhancement Ratio (RER) was calculated for ONC201 (5, 10 µM) and Radiation (2, 4 and 8 Gy) out of 3 independent experiments. (c) Cell death analysis through Annaxin V/PI detection of HT-29 cells treated with ONC201 (5 and 10 µM) 24 h prior Radiation. Samples were harvested at 72 h. One-way ANOVA test has been run comparing treated vs Vehicle and mock radiated on n = 3 experiments, *p ≤ 0.05; **p < 0.01; ***p < 0.001 (± SD).
