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Fig. 6. Overexpression of SMAD6 in ES cells inhibits the generation of FLK1+ cells. (A) ES cells stably expressing SMAD6 were differentiated and subjected to FACS analysis for FLK1 expression. Numbers in insets indicate the percentage of FLK1+ cells. 2Ab indicates cells stained with secondary antibody alone. Flag-6 and Flag-8 indicate control ES cell clones. Flag SMAD6-6 and SMAD6-7 represent two independent ES cell clones stably expressing Flag-SMAD6. Expression of exogenous Flag-SMAD6 was verified with anti-FLAG antibody (arrow). (B) Results from four independent experiments are shown as a percentage of Flag-6 control. Error bars indicate s.e.m.