Extended Data Fig. 1: Control experiments demonstrating NDF-FACT interactions in vitro and their synergistic stimulation of transcription.
From: Phase-separated NDF−FACT condensates facilitate transcription elongation on chromatin
a, SDS-PAGE analysis of FACT purified from Sf9 cells, with proteins separated on an 8% SDS-PAGE gel and visualized using Coomassie Blue staining. Three biological replicates were performed. b, Analysis of the interaction between recombinant NDF-his (0.3 µM) and FACT-FLAG (0.15 µM) using a Ni-NTA pull-down assay, with products separated on a 12% SDS-PAGE and detected by silver staining. Three biological replicates were performed. c, Nucleosome transcription assay demonstrates the synergistic effect between 0.2 µM NDF and 0.2 µM FACT when added either sequentially with 10-minute intervals or premixed before introduction to the Pol II elongation complex before adding 1 mM NTPs. The graph on the right shows statistical analysis of the run-off products based on three independent experiments (mean ± SD, two-tailed t-test).
