Transient Downregulation of Nanog and Oct4 Induced by DETA/NO Exposure in Mouse Embryonic Stem Cells Leads to Mesodermal/Endodermal Lineage Differentiation

Figure 1

Reexpression of pluripotency genes following exposure to DETA/NO. (a) Western blot analysis of Nanog and Oct4 proteins. D3 mESCs were grown in the presence (+LIF) or in the absence (−LIF) of LIF for 3 days. Cells were then cultured as indicated in Section 2 for additional periods of 1 and 2 days (day 4 and day 5). C: control cells. T: cells exposed to 1 mM DETA/NO for 19 h on day 4. Lanes 5 and 6 refer to cells exposed to DETA/NO and subsequently cultured for 1 and 2 days as in Section 2. (b) GFP expression in D3-Oct4 mESCs. Control cells were cultured in the presence of LIF for 6 days (+LIF/Control). Treated cells were cultured in the presence of 1 mM DETA/NO for 19 hours on day 4 and subsequently cultured as indicated in Section 2 for an additional period of 2 days in absence of LIF. PhC: phase contrast. Results are representative of 3 independent experiments. (c) Western blot analysis reveals Nanog and Oct4 reexpression after DETA/NO treatment. +/− LIF indicates that cells were cultured for 4 days in the presence of LIF, then LIF was removed and cells were subsequently exposed to 1 mM DETA/NO for 19 hours. Days 5 and 6 indicate that cells were subsequently cultured for 1 and 2 days in the absence of LIF. −/+LIF indicates cells that were cultured for 4 days in the absence of LIF; exposure to 1 mM DETA/NO for 19 hours was then performed in the presence of LIF. Days 5 and 6 indicate that cells were subsequently cultured for additional periods of 1 and 2 days, respectively, in the presence of LIF. Results are representative of 3 independent experiments.

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