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. 2015 Nov 13;5:16595. doi: 10.1038/srep16595

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Copyright © 2015, Macmillan Publishers Limited

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(a) Schematic illustration depicting the reporter design. CRISPR-Cas9 was used to target the stop codon of BRN3B in H7 hESCs. A P2A linked membrane targeted mCherry was added to the BRN3B ORF by homologous recombination. Following translation, the BRN3B transcription factor protein is localized to the nucleus while mCherry is targeted to the cell membrane. (b) Schematic of RGC differentiation protocol. (c) Phase microscopy of differentiation over time. Cultures become confluent by day 10. (d) Phase and fluorescence comparison microscopy of differentiation. mCherry fluorescence becomes visible on day 25. More mCherry+ cells form over time and cell bodies become clearly visible. (c,d) Scale bars = 100 μm.