Fig. 3: Branaplam lowers HTT-transcript and protein levels in normal and HD patient cells.

a Quantitative PCR results showing the transcript levels of HTT exon 50a included after DMSO or branaplam treatment (at indicated doses), ****P < 0.0001, one-way ANOVA. The data are presented as mean ± SEM of three independent experiments. b HTT-transcript changes after DMSO or branaplam treatment (at indicated doses) for 24 h in SH-SY5Y human neuroblastoma cells, ***P < 0.001, ****P < 0.0001, one-way ANOVA. The data are presented as mean ± SEM of three independent experiments. c Western blot and d quantitation showing total HTT protein levels after DMSO or branaplam treatment (at indicated doses) for 48 h in SH-SY5Y cells, *P < 0.05, ***P < 0.001, ****P < 0.0001, one-way ANOVA. The data are presented as mean ± SEM of three independent experiments. e Quantitative PCR results showing HTT mRNA expression levels after DMSO or Branaplam treatment (at indicated doses) for 24 h. Two independent HD patient fibroblasts cell lines (GM04723 and ND31551 (L5)), and a TaqMan™ assay priming for human HTT exons 64–65 was used. Samples are normalized to human GAPDH, and data are the mean ± SEM relative to HD fibroblasts treated with DMSO (eight biological replicates, *P value <0.001, one-way ANOVA followed by a Bonferroni’s post hoc). f Quantitative PCR results showing HTT50a exon inclusion levels in two HD patient fibroblasts cell lines (GM04723 and ND31551 (L5)) after 24-h treatment with DMSO or branaplam (at indicated doses). Samples are normalized to human GAPDH, and data are the mean ± SEM relative to HD fibroblasts treated with DMSO (four biological replicates, *P value <0.001, one-way ANOVA followed by a Bonferroni’s post hoc). g western blot and h, i quantitation showing mutant HTT protein levels after DMSO or branaplam treatment (at indicated doses) for 96 h in HD patient fibroblast cells, ****P < 0.0001, one-way ANOVA. Data are the mean ± SD relative to DMSO from four independent experiments.