Extended Data Fig. 1: Molecular aspects of p16 activation during in vitro and in vivo reprogramming.
From: p16High senescence restricts cellular plasticity during somatic cell reprogramming
a) Colocalization of p16High (EGFP +) cells with markers of DNA damage response (pATM, pKu80, pRad51) and cell cycle inhibitors p27 and p57. Positive controls for pATM, pKu80, pRad51, p27 derived by treatment DF with doxorubicin (500 nM, 24 h). Mouse placenta serves as a positive control for p57. Scale bars – 50 μm. b) Teratomas emerged during in vivo reprogramming in p16Cre;i4F;rtTA;mTmG mice produce all three germ layers. H&E staining shows the presence of Car, cartilage (mesoderm); ET, endodermal tube (endoderm); NR, neural rosette (ectoderm). Scale bar represents 100 μm. c) Immunofluorescent analysis of teratomas, obtained by in vivo reprogramming of p16Cre;i4F;rtTA;mTmG mouse co-stained for p16High and mesenchymal (Desmin, a-SMA), endothelial (Cd31), macrophagic (F4/80) and ectodermal (GFAP) markers. Scale bars – 50 μm. d) Immunofluorescent analysis of teratomas derived by grafting feeder-free p16Cre;i4F;rtTA;mTmg iPS cells into NSG mouse for p16High cells and mesenchymal markers Desmin and α-SMA, vascular endothelial marker Cd31, macrophage marker F4/80 and neuroectodermal marker Gfap. Scale bar – 50 μm.
