1. Myc‐induced miRNAs attenuate differentiation of c‐Myc‐depleted ES cells on LIF withdrawal.
Myc‐induced miRNAs attenuate differentiation of c‐Myc‐depleted ES cells on LIF withdrawal. (A) The percentage of cells staining positive for SSEA‐1 at the indicated time points was quantitated by flow cytometry after removal of LIF from mES cells. Before LIF withdrawal mES cells were infected with empty lentiviral vector alone (red bars); with lentiviral vector expressing c‐myc (blue bars); or lentiviral vector expressing shRNA against c‐myc (green bars). Data are expressed as the percentage of SSEA‐1‐positive mES cells in the absence of LIF relative to mES cells in the presence of LIF. *P‐value<0.05, **P‐value<0.01, ***P‐value < (B) mES cells were infected with empty lentiviral vector (WT) or lentiviral vector expressing shRNA against c‐myc. After selection, the c‐Myc knockdown ES cells (see A above) were either electroporated with a mixture of miRNAs (miR‐141, ‐200, ‐338, ‐429) or electroporated without addition of miRNAs. Twenty‐four hours after transfection, LIF was withdrawn from the medium. Left panel shows WT ES cells maintained in LIF for the time course of the experiment. A single well is shown for each time point. (C) SSEA‐1 flow cytometry 7 days after LIF withdrawal from c‐myc knockdown ES cells transfected as indicated in the inset. *P‐value<0.05, **P‐value<0.01, ***P‐value <0.001.
Chin‐Hsing Lin et al. EMBO J. 2009;28:
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