Figure 5.

PIN1 regulates mHTT half-life. HEK293 cells were co-transfected with htt_1–171Q60GFP and HA-PIN1 or HA-PIN1DM. Cells were harvested for analysis at indicated time points. (A) Representative western blot showing htt_1–171Q60GFP upon MG-132 treatment (10 μM for 6 h) in the presence of PIN1 as compared to PIN1DM. Western blot also shows PIN1, CYCLIN-D1, and β-ACTIN as loading control. Cells were treated 24 h after transfection. (B) Representative western blot showing htt_1–171Q60GFP levels upon treatment with CHX (40 μg/ml), at different time points, in the presence of PIN1 as compared to PIN1DM. Westerm Blot also shows PIN1, CYCLIN-D1, and β-ACTIN as loading control. CHX treatment started 6 h after transfection. (C) Relative htt_1–171Q60AGFP protein level based on western blots quantification as shown in (B). Data are the mean ± SEM from 6 independent experiments using 2 different batches of cells. ^*P < 0.05, ^***P < 0.001. Asterisk indicates the statistically significant difference in the level of protein between PIN1 and PIN1DM-expressing cells at the indicated time point. Values were calculated relative to corresponding t0 sample.