Aqueous Extract of Paeonia lactiflora and Paeoniflorin as Aggregation Reducers Targeting Chaperones in Cell Models of Spinocerebellar Ataxia 3
High-content compound screen using Flp-In 293 cells with inducible ATXN3/Q75-GFP expression. (a) Experiment flow chart. ATXN3/Q75-GFP 293 cells were plated into 96-well dishes, grown for 24 hr and treated with different concentrations of the herb or compound for 8 hr. Then doxycycline and oxaliplatin was added to the medium to for 6 days and aggregation percentage was assessed by HCA system. (b) Representative fluorescence microscopy images of ATXN3/Q75-GFP cells untreated or treated with P. lactiflora (10 µg/mL) or paeoniflorin (100 nM) for 6 days. (c) Aggregation analysis of ATXN3/Q75-GFP cells untreated or treated with aqueous extract of P. lactiflora (2~200 µg/mL), paeoniflorin, garlic acid, albiflorin and SAHA (100 nM~5 µM). To normalize, the relative aggregation level in untreated cells is set as 100%. The red line represents 85% aggregation with SAHA treatment (100 nM).