Fig. 6: Atoh8 constrains cellular plasticity during reprogramming and transdifferentiation.
a, Experimental set-up. b, Immunofluorescence of Pou5f1-GFP+ colonies at day 15 of reprogramming. Scale bars, 100 µm. c, Pou5f1+ colony quantification (n = 3 independent experiments). d, Histological analysis of teratomas. Two independent teratomas were analysed per cell line. Scale bars, 1 mm. e, Graph depicting the percentage of Pou5f1-GFP+ cells (n = 3 independent experiments). f, FACS analysis showing the emergence of Pou5f1-GFP+ cells. g, Experimental design. h, Image representing AP+ colonies at day 15 of reprogramming, representative of three independent experiments. i, Colony quantification (n = 3 independent experiments). j, Western blot for Sox2, Ssea1, Nanog, Oct4 and Actin in Control and Atoh8 KD-derived iPS cell lines. k, Schematic of human induced pluripotent stem cell reprogramming. l, Picture representing SSEA4+ colonies at day 26 of reprogramming, representative of two independent experiments. Scale bars, 100 µm. m, Colony quantification (n = 2 independent experiments). n, Immunofluorescence for Oct4 and Nanog. hiPS, human induced pluripotent stem. Scale bars, 100 µm. o, Schematic depicting MEF-to-neuron transdifferentiation. p, MAP2 immunostaining. Scale bars, 100 µm. q, MAP2+ cell quantification per field (n = 2 independent experiments). In c, e and i, the data represent means ± s.d. and statistical significance was determined by two-tailed Student’s t-test.
