Fig. 6

Hypoxia-mediated reprogramming to pluripotent stem cells is dependent on PLD2. (A) Representative images of induced pluripotent stem cells (iPSCs) by reprogramming mouse embryonic fibroblasts (MEFs) with OSKM genes (Yamanaka factors Oct3/4, Sox2, Klf4 and cMyc) and Nanog reporter retroviruses. Top, bright field microscopy of iPSCs. Medium, immunofluorescence image showing GFP in cells in which Nanog is active. Bottom, Alkaline phosphatase activity. (B) Left, quantification of iPSCs generated from MEFs infected with OSKM genes and an additional lentiviral vector that expresses GFP in cells in which the Nanog promoter/enhancer is active and the corresponding plasmid overexpressing PLD2, shPLD2, both or carrying Ev under normoxia or hypoxia. Right, quantification of cells with alkaline phosphatase activity at the end of the iPSC generation experiment. A minimum of three biological were performed per each experiment. The data were analysed using Student’s t test. Asterisks indicate statistical significance with respect to Ev carrying cells, unless indicated by horizontal lines. *, P < 0.05; **, P < 0.01; ***, P < 0.001