Extended Data Fig. 2: ZFP462 is conserved across vertebrates and acts as transcriptional repressor.

a) Phylogenetic tree of ZFP462 protein orthologues in vertebrate species. Bootstrap values are shown on branches. b) Western blot shows expression ZFP462 fusions with TetR-FLAG in CiA Oct4 dual reporter ESCs. Bottom image shows long exposure. Hashtag indicates cleaved TetR-FLAG protein from TetR-FLAG-ZFP462-FL (full-length). TetR-FLAG fusions initially express mCherry for selection which is cleaved via P2A signal. Asterisk marks P2A uncleaved protein product. c) Representative flow cytometry histograms show GFP expression in CiA Oct4 dual reporter ESCs expressing TetR-FLAG fusions with full-length or truncated ZFP462 proteins. Each histogram is average profile of 100,000 analysed cells.