Fig 1.

RYBP-deficient ES cells. (A and B) Western blot of cell extracts and phase-contrast images from cultures after 4 days of ethanol (EtOH) or hydroxytamoxifen (4′-OHT) treatment. (C) BrdU incorporation during a 20-min pulse in cells grown for different times after treatments (4 or 12 days [+d4 or +d12, respectively]). (D) Expression of stem cell markers in ethanol-treated (wild-type) or 4′-OHT-treated (RYBP-KO) ES cells identified by immunofluorescence (Oct4 and SSEA1) or immunohistochemistry (alkaline phosphatase [AP]). RYBP immunofluorescence was included as a control of gene inactivation. (E) Deregulated differentiation of RYBP-mutant ES cells. Time course (days in x axis) gene expression analysis by RT-qPCR of mRNAs encoding cell lineage markers; error bars are SDs of relative mRNA levels indicated in the y axis.