Myeloid Zinc Finger 1 (Mzf1) Differentially Modulates Murine Cardiogenesis by Interacting with an Nkx2.5 Cardiac Enhancer
Figure 5
In vitro differentiation of the double-transgenic dox-inducible Mzf1 overexpressing tetOMzf1-Nkx2.5 CE eGFP ES cell line.
Scale bars: 200 µm for all panels. A. Lentiviral constructs for the production of the doxycyclin-inducible tetOMzf1-Nkx2.5 CE eGFP ES line. LTR: long terminal repeats. TRE: tetracyclin responding element. CMV: cytomegalovirus promoter. IRES: internal ribosomal entry site. rtTA: reverse tetracyclin transactivator. B. In vitro differentiation protocols with time-schedules of dox-treatment. C. Morphology of differentiating EBs on day eight. Permanent and day 0–5 dox-treatment led to closely packed globular clusters. In contrast dox-treatment from day 5 showed a normal differentiation pattern comparable to the control w/o dox. D/E. FACS analysis revealed a significant increase in eGFP+ CPCs for dox-treatment from day 5 of differentiation whereas a continuous and day 0-5 dox-treatment resulted in a significant decrease of eGFP+ CPCs compared to control w/o dox; ** = p <0.01.