Fig 1: Immunoprecipitation of wtHtt and mHtt interacting proteins. A) SDS-PAGE western blot (WB) of input, unbound fraction and immunoprecipitation of quadruplicate control (Htt-Q20), wild-type Htt (Flag-Htt-Q20) and mutant Htt (Flag-Htt-Q140) samples. WB was performed using antibodies directed against the Htt N-terminus, Flag-tag, and an antibody directed against vinculin was used as a negative control. B) Volcano plots showing proteins significantly up in Q20 vs. control and Q140 vs. control. The proteins that pass the p-value and log2 fold change threshold are shown in green. The tables for Q-length groups vs. control are given in the Supplementary Table 2. C) Venn diagram showing the overlap between the proteins significantly up in Q20 vs. control and Q140 vs. control. D) SDS-PAGE WB was performed to validate the results obtained by mass spectrometry. Specific validation of Flag-tagged wtHtt and mHtt was done using D7F7 antibody for Htt and a-Flag antibody. Validation of wtHtt and mHtt interactor F8a1/Hap40 and mHtt interactor Ahsa1 are shown. Vinculin was used as a negative control. E) Biological processes enriched in Q20 and Q140. p-value indicated by color, frequency of GO-term indicated by node size, degree of similarity indicated by thickness of edges.