Extended Data Fig. 4: Potential influence on c-Raf and KSP pathways following treatment with the mHTT–LC3 linker compounds.
From: Allele-selective lowering of mutant HTT protein by HTT–LC3 linker compounds
a, Representative results (from three biological repeats) of the in vitro c-Raf kinase assay (see Methods) showing that only 10O5 inhibits c-Raf activity within the concentration range tested. b, Representative western blots and quantifications of phospho-MEK and phospho-ERK as indicators of Raf activity (left) and phospho-BUBR1 as an indicator of KSP inhibition (right) in cultured cortical neurons treated with indicated compounds (100 nM for 10O5, 8F20, AN1, and 50 nM for AN2) or the DMSO control. c, Similar to b, but in immortalized fibroblasts from a patient with HD (Q47). Note that phospho-BUBR1 is essentially absent and too weak to quantify, indicating that KSP was not inhibited by any of the compounds at the concentration tested. Data are mean ± s.e.m. In b, c, all data were corrected by the loading control (β-tubulin) and normalized to the averaged signal of the DMSO control group. The statistical analysis was performed by one-way ANOVA and F, degree of freedom and post hoc P values are indicated in each bar plot. The n number indicates the number of independently plated and treated wells.
