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. Author manuscript; available in PMC: 2016 Feb 22.
Published in final edited form as: Nat Commun. 2014 Jun 17;5:4112. doi: 10.1038/ncomms5112

Figure 1. Direct conversion of human fibroblasts into neuronal cells by p16 and p19 shRNAs.

Figure 1

(a) Schematic drawing of the structure of the Ink4a/Arf locus and the experimental strategy to derive induced neural cells. The images show the IMR90 fibroblast morphology and TUJ1-positive iN cells (shp16) with multiple neurites processing out of the cell body 4 weeks after induction. (b) Depletion of p16, p19 and p16/p19 by shRNAs. Western blot analysis shows decrease of p16 and p19 expression by shRNAs in IMR90 cells. (c) Characterization of neuronal cells derived from shp16, shp19 and shp16/19 IMR90 cells. The images show the TUJ1- and MAP2- positive iN cells from shp16, shp19 and shp16/19 IMR90 fibroblasts 3 weeks after induction. Scale bar, 10 μm. (d and e) Quantitative analysis of iN cells from IMR90 cells with or without expressing shp16, shp19 or empty vector after induction. Kinetic analysis of iN purity was counted by morphology, TUJ1- (early stage, before one week induction) and MAP2-positive (late stage, after 1 week induction) staining, and the numbers represent the percentage of iN cells at the time point of quantification (d). Neuronal yield was determined as the percentage of MAP2+ cells in relation to the initial number of plated fibroblasts at 3 week after induction (e). Quantitative data were mean±s.e.m. from at least three independent experiments.