A and B, Flies expressing PrP ubiquitously (da>PrP-M9) were fed with combinations of 17-DMAG at 0, 48, and 96 µg/ml and dexamethasone at 0, 24, and 48 µg/ml during development. Analysis of PrP levels in 1 day-old flies by western blot showed significant reduction in PrP in two combinations following quantification and analysis by ANOVA, with 17-DMAG 96 µg/ml and dexamethasone 24 µg/ml inducing the strongest response. C, Analysis of PrP and GFP mRNA by qPCR showed no significant changes in flies treated with the optimum drug combination. D and E, Western blot and quantification of Hsp70 in flies treated with different combinations of 17-DMAG and dexamethasone. Quantification of Hsp70 followed by normalization with a non-specific band (control) revealed that all treatments induced significant increases in Hsp70, with 17-DMAG 96 µg/ml and dexamethasone 24 µg/ml inducing the strongest response. F, Expression of Hsp70 mRNAs by qPCR revealed a four-fold increase in flies treated with the optimum 17-DMAG/dexamethasone cocktail in flies expressing either PrP or CD8-GFP. G, Analysis of Hsc70-4 and Hsp27 mRNA by qPCR in flies treated with the optimum 17-DMAG/dexamethasone cocktail showed no significant changes in these two chaperones. Statistical significance was analyzed by ANOVA multiple comparisons for western blots and t-test for qPCR: * p<0.05; ** p<0.01; *** p<0.001.