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. Author manuscript; available in PMC: 2010 Aug 5.
Published in final edited form as: Differentiation. 2010 May 27;80(1):9–19. doi: 10.1016/j.diff.2010.05.001

Fig. 1.

Fig. 1

Loss of c- and N-myc disrupts mESC growth and triggers lineage commitment. (A) Schematic of the floxed c- and N-myc loci. (B) Detection of c- and N-myc gene copy number changes by real-time qPCR. Gene copy numbers for c- and N-myc were normalized using two reference genes, β-actin and Nat1. Error bars are standard deviations. Decreases in c- and N-myc copy numbers had p values of 8.3×10−7 and 7.2×10−7, respectively, in cDKO2, and p values of 1.6×10−7 and 2.4×10−5 in cDKO3, respectively. (C) Phase contrast image of mESC lines in the presence or absence of CRE expression. Arrow denotes differentiated colony. GFP and GFP-Cre denoted cell cultures transduced with MSCV ires-GFP and MSCV Cre-ires-GFP retroviruses, respectively. (D) Real-time qRT-PCR of c-myc, N-myc, and a series of different lineage-specific marker genes in WT and DKO mESC lines. Levels of specific mRNAs measured by qRT-PCR were normalized to the levels of the loading control Nat1. Error bars are standard deviations. Changes in relative expression of N-myc, c-myc, Ngn3, Gata6, Bmp4, and Fgf5 had p values of 7×10−4, 2×10−4, 1×10−3, 8.6×10−5, 9×10−4, and 9.6×10−4, respectively, in cDKO2 and p values of 8.7×10−5, 1.4×10−5, 2.7×10−4, 3.4×10−4, 5.1×10−4, and 1.3×10−3, respectively, in cDKO3. (E) RT-PCR of hematopoietic, neural and sensory organ differentiation markers in mESC of the indicated genotype.