Fig. 3

J3 reduces mHTT. A HeLa cells stably expressing CFP-103Q were treated with 20 μM of J3 for 48 h, the mHTT (disperse cyan fluorescence or aggregates) spots were counted from 100 cells, and quantified. Scale bar = 20 μm. B CFP-103Q-HeLa cells were treated with 20 μM of J3 for 12, 24, 48 h, respectively. The insoluble (I-mHTT) and soluble mHTT (S-mHTT) were detected by anti-GFP (sc-9996) for western blot. C–D The quantification of insoluble (C) and soluble mHTT (D) at different times was analyzed. E CFP-103Q-HeLa cells were treated with 5, 10, and 20 μM of J3 for 48 h, the protein level of mHTT (anti-GFP, sc-9996) were measured by western blot. F–G The quantification of insoluble (F) and soluble mHTT (G) treated by different doses of J3 was analyzed. H WT-HeLa cells were treated with 20 μM of J3 or 1 μM of Rap for 48 h, the endogenous HTT (anti-HTT, MAB2166) was measured by western blot. I The protein level of endogenous HTT of H was quantified. Data are presented as mean ± sem from three individual experiments or three different fields. *p < 0.05, **p < 0.01, ***p < 0.001, ns means not significant