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. 2022 Nov 10;13:6797. doi: 10.1038/s41467-022-34419-x

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Paradigm: cell types and parameters analyzed. Created with BioRender.com. b–d tHTT levels (2B7/D7F7 assay), mHTT levels (2B7/MW1 assay) and toxicity (adenylate kinase release, Triton: positive control) in fibroblasts (b), iPSC (c) and cortical progenitors (d) with different Branaplam concentrations for 72 h. Green: Ctrl samples (n = 2 33Q/33Q-S1-109, 4L6/4L6-S1-027), purple: HD samples (n = 2 919/919-S1-101, 4Q4/4Q4-S1-109). b Fibroblasts statistics: tHTT: one-way ANOVA with Geisser–Greenhouse correction (P = 0.0012); mHTT: no statistics applied; toxicity (Triton excluded): Friedman test (P = 0.0770). c iPSC statistics: tHTT one-way ANOVA with Geisser–Greenhouse correction (P = 0.0032); mHTT: no statistics applied; toxicity (Triton excluded): one-way ANOVA with Geisser–Greenhouse correction (P = 0.0455), no significant differences in multiple comparisons. d cortical progenitor statistics: tHTT IC₅₀: 919-S1-101 = 2.233 × 10⁻⁹M; 4Q4-S1-109 = 6.102 × 10⁻⁹M; 33Q-S1-109 = 3.191 × 10⁻⁹M; 4L6-S1-027 = 1.182 × 10⁻⁹M; mHTT: 919-S1-101 = 5.528 × 10⁻⁹M; 4Q4-S1-109 = 8.952 × 10⁻⁹M; 33Q-S1-109 and 4L6-S1-027 = not calculated; toxicity (Triton excluded): one-way ANOVA with Geisser–Greenhouse correction (P = 0.06). e tHTT levels (2B7/D7F7 assay), mHTT levels (2B7/MW1 assay) (Ctrl n = 3; HD n = 4) and toxicity (adenylate kinase release, Triton as positive control) in cortical neurons of (Ctrl n = 4; HD n = 4). tHTT and mHTT measured with DMSO (dark shades) or 10 nM Branaplam (light shades) for 72 h. Statistics: tHTT: 2-way ANOVA (DMSO vs. Branaplam: P = 0.0009; Ctrl vs. HD: P = 0.6820; interaction: P = 0.3833); mHTT: two-way ANOVA (DMSO vs. Branaplam: P = 0.0614; Ctrl vs. HD: P = 0.0223; interaction: P = 0.0615); toxicity (Triton excluded): Friedman test (P = 0.5306). Bars: median ± IQR. f Bar plot showing number of Casp3/7 positive beta-III-Tubulin+ and CTIP2+ cortical neurons after DMSO (dark shades) or (light shades) 72 h 10 nM Branaplam treatment (Ctrl n = 4; HD n = 4). Statistics: beta-III-Tubulin+: two-way ANOVA (DMSO vs. Branaplam: P = 0.0782; Ctrl vs. HD: P = 0.4744; interaction: P = 0.9502); CTIP2+: 2-way ANOVA (DMSO vs. Branaplam: P = 0.7230; Ctrl vs. HD: P = 0.5779; interaction: P = 0.6348). Bars: median ± IQR. Source data are provided as a Source Data file.