Fig. 1
From: LSD1 is required for euchromatic origin firing and replication timing
LSD1 interacts with the DNA replication machinery. a Immunoaffinity purification of LSD1-containing complexes. HeLa cells stably expressing FLAG-LSD1 were synchronized by double-thymidine block. Cellular extracts were immunopurified with anti-FLAG affinity columns and eluted with FLAG peptides. The eluates were resolved by SDS-PAGE and visualized by silver staining. The proteins bands were retrieved and analyzed by mass spectrometry (left). Column-bound proteins were analyzed by western blotting using antibodies against the indicated proteins (right). The percentage of input was 5%, the same for the experiments described below. b Immunoprecipitation of endogenous proteins in S-phase HeLa and U2OS cells with antibodies against LSD1, followed by immunoblotting with antibodies against the indicated proteins. c Schematic representation of the structure of LSD1. d GST pull-down assays with GST-LSD1, GST-N (1-171 aa), GST-SWIRM (171-271 aa), and GST-Tower (416-521 aa) and in vitro transcribed/translated MCM2-7. Coomassie brilliant blue (CBB) staining of GST-fused wild-type and deletion mutants of LSD1 is shown (arrows)
