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. Author manuscript; available in PMC: 2018 Sep 14.
Published in final edited form as: Cell Rep. 2018 Aug 14;24(7):1852–1864. doi: 10.1016/j.celrep.2018.07.046

Figure 2. Maf1 Promotes the Expression of Mesodermal Markers in mEBs.

Figure 2.

(A) qRT-PCR analysis of markers associated with the three germ layers (endoderm: GATA6 and GATA4; mesoderm: T and Mesp1; ectoderm: Nestin and SOX1) in control and Maf1-knockdown mESCs and mEBs at the indicated days after cells were infected with either a control lentiviral construct or one harboring an shRNA-targeting Maf1 mRNA.

(B) qRT-PCR analysis of Maf1, T, and Oct4 in control and Maf1-induced mESCs and mEBs at day 3. mESCs were differentiated into EBs after infection with either a control lentiviral construct (rtTA) or one expressing an HA epitope-tagged Maf1 construct (Maf1-HA). Control and Maf-HA cells were treated with 1 μg/mL doxycycline (dox) 2 days before formation of mEBs and throughout the process of mEB formation. In (A) and (B), data are mean ± SD of n = 3 independent experiments. Transcript amounts were normalized to β-actin, and the fold change was calculated relative to the amount of transcript in control mESCs. *p < 0.05, unpaired Student’s t test.

(C) Immunoblot analysis of Maf1-HA, T, and β-actin protein expression in mESCs and mEBs at the indicated days. Protein amounts were normalized to β-actin, and the fold change was calculated relative to the amount of protein in control mESCs.