hygromycin invivogen 293 atxn3 q 75 gfp cells were plated into 96 well 2 u00d7 10 4 well dishes grown for 24 hr and treated with different concentrations of the trehalose or analogs 100 nm u02dc 100 u03bcm and suberoylanilide hydroxamic acid saha cayman chemical for 8 hr then doxycycline 10 u03bcg ml bd was added to the medium in each well to induce atxn3 q 75 gfp expression for 6 days oxaliplatin 5 u03bcm sigma was also added to increase aggregate accumulation through inhibition of cell division then cells were stained with hoechst 33342 0 1 u03bcg ml sigma and aggregation percentage was assessed by high content analysis hca system imagexpressmicro molecular devices with excitation emission wavelengths at 482 536 egfp sh sy5y tbp q 79 gfp cells were seeded in 6 well 2 u00d7 10 5 well plate with all trans retinoic acid 10 u03bcm sigma added at seeding time at day 2 cells were treated with trehalose or analogs 10 u03bcm for 8 hr then doxycycline 5 u03bcg ml was added and the cells were kept in the medium containing 10 u03bcm trans retinoic acid doxycycline and trehalose analogs for 7 days after that cells were stained with hoechst 33342 0 1 u03bcg ml and aggregation percentage was assessed by hca hydrolysis of trehalose and analogs was tested using porcine kidney trehalase sigma aldrich t8778 the standard reaction was performed by adding 0 001 unit of trehalase into 50 u03bcl reaction solution containing 135 mm citric acid ph 5 7 and 28 mm trehalose or trehalose analogs in addition the inhibition of analog against the trehalase activity was also examined the standard reaction was performed by adding.