Extended Data Fig. 1: Validation of XL-MS for protein interaction analysis.
From: A chaperone-like function of FUS ensures TAZ condensate dynamics and transcriptional activation
a, Schematic depicting the chemical synthesis procedure of L-NHSF. b, Purified recombinant proteins were analyzed by SDS-PAGE and visualized by Coomassie blue staining. c, Purified PRFA and PRMC were untreated or cross-linked with NHSF or L-NHSF and analyzed by Western blotting (WB). d & e, PRFA and PRMC cross-linked with NHSF or L-NHSF were analyzed by mass spectrometry to identify the number of cross-linked peptides (d) and types of cross-linking sites (e). f, Distribution of Cα-Cα distance of cross-linked residues from PRFA and PRMC samples treated with NHSF or L-NHSF. g, GFP-T1-IDR was cross-linked with the indicated concentrations of L-NHSF at 37.5 mM or 150 mM NaCl and analyzed for the formation of droplets. Right: Quantification of GFP-T1-IDR phase separation in the presence of different concentrations of L-NHSF. Data are mean ± SEM. n = 3 biologically independent samples. h, Schematic depicting the crosslinking of GFP-T1-IDR at the indicated NaCl concentrations and then followed by mass spectrometry analysis. i, GFP-TAZ was cross-linked with the indicated concentrations of L-NHSF at 37.5 mM or 150 mM NaCl and analyzed as in g. Bottom: Quantification of the sizes of droplets in the presence of different concentrations of L-NHSF. n = 500 droplets in each group. j, Hela cells expressing WW-F or WW-F plus WW-Strep were subjected to co-immunoprecipitation analysis. Whole cell extract (WCE) and Streptactin immunoprecipitates (IPs) were analyzed by WB. For d, e, f, and i, data are representative of three independent experiments, with similar results obtained.
