Fig. 2: Characterization of the roadblock gene KO in different reprogramming systems and kinetics. | Nature Communications

Fig. 2: Characterization of the roadblock gene KO in different reprogramming systems and kinetics.

From: B1 SINE-binding ZFP266 impedes mouse iPSC generation through suppression of chromatin opening mediated by reprogramming factors

Fig. 2

A, C Cas9 expressing Nanog-GFP MEF reprogramming with MKOS (A), STEMCCA (C) piggyBac transposons with sgRNA expression at day 15. Red; mOrange, Green; Nanog-GFP. B, D Nanog-GFP+ and Nanog-GFP- mean colony numbers from A and C. The graphs in B and D represent an average of three independent experiments. E Cas9 expressing NSC reprogramming with MKOS piggyBac transposons with sgRNA expression at day 15. Green; immunofluorescence for NANOG. F NANOG + and NANOG- mean colony numbers from E. The graph represents an average of two independent experiments. G Accelerated CD44/ICAM/Nanog-GFP expression changes by sgRNA expression against the roadblock genes (n = 2). Red; Nanog-GFP- cells, Green; Nanog-GFP + cells. H Quantification of Nanog-GFP + cells from day 6 to 10 of reprogramming. The graph represents an average of two independent experiments. In B, D, F, ****p < 0.0001, ***p < 0.001, **p < 0.01,*p < 0.05. Source data and exact p-values are provided as a Source Data file.

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