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. 2021 Sep 6;12:5305. doi: 10.1038/s41467-021-25651-y

Fig. 6. CHIR99021 treatment stabilizes CAST in HD models.

Fig. 6

Total lysates from HdhQ7 and HdhQ111 cells were harvested after the indicated treatments; 3 μM CHIR99021 for 48 h followed by a 2.5 μM lactacystin (Lac) for 12 h or b 250 nM epoxomicin (Epo) for 12 h and subjected to western blotting with the indicated antibodies. Histograms show the relative abundance of CAST (n = 4), calpain I (n = 3), or MCL1 (n = 4). c Calpain enzymatic activity was examined in HdhQ7 and HdhQ111 cells with the indicated treatments: 48 h treatment with CHIR99021 followed by 12 h treatment with Lac (n = 4) or Epo (n = 4). d Total immunoprecipitation (IP) with an anti-CAST antibody followed by western blotting with an anti-ubiquitin antibody. Representative immunoblots and quantification of the relative abundance of ubiquitinated CAST (n = 3). All values are reported as mean ± SEM. Data are representative of at least three independent experiments. Data were compared using one-way ANOVA with Tukey’s post hoc test. Exact p values are shown in the figures.