Fig. 3
From: ZNF516 suppresses EGFR by targeting the CtBP/LSD1/CoREST complex to chromatin
ZNF516 interacts with CtBP/LSD1/CoREST complex in vivo and in vitro. a Co-fractionation of ZNF516 and CtBP/LSD1/CoREST complex by FPLC. MCF-7 cell proteins were extracted, concentrated, and then fractionated on Superose 6 size exclusion columns. Chromatographic elution profiles and immunoblotting analysis of the chromatographic fractions are shown. The elution positions of calibration proteins with known molecular masses are indicated, and an equal volume from each fraction was analyzed. b Silver staining and western blot analysis of the ZNF516-containing complex fractionated by Superose 6 gel filtration. Cellular extracts from HEK293T cells transfected with FLAG-ZNF516 were fractionated on Superose 6 size exclusion columns. Chromatographic elution profiles, silver staining, and western blot analysis of the chromatographic fractions are shown. The elution positions of calibration proteins with known molecular masses are indicated, and an equal volume from each fraction was analyzed. c Schematic diagrams of ZNF516, CtBP1/2, and CoREST deletion mutants. d GST pull-down experiments were performed with bacterially expressed GST or GST-fused ZNF516 deletion mutants and in vitro transcribed/translated truncation mutants of CtBP1, CtBP2, or CoREST, or full length of LSD1, HDAC1 or HDAC2. Coomassie brilliant blue staining of the GST-fused proteins was shown with arrows. e Bacterially expressed GST or GST-fused indicated proteins were performed GST pull-down experiments with in vitro transcribed/translated ZNF516
