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. 2019 Jan 30;33(4):5287–5299. doi: 10.1096/fj.201801110RR

Figure 1.

Figure 1

Rapid differentiation of human iNs induced by overexpression of NEUROG2/1 in hESCs. A) Schematic outline of the lentiviral constructs and flow diagram of the differentiation protocol. B) Representative phase-contrast images illustrating the changes in cell morphology on d 1, 2, and 4 after doxycyclin (Dox) induction. Scale bar, 100 μm. C) Representative immunofluorescence images of hESCs and iN cells immunostained with the stem cell markers Nanog and Oct3/4. One hundred percent of iNs lost staining for both Nanog and Oct3/4 on d 4. Scale bar, 100 μm. D) Representative images of hESCs and iN cells immunostained for the neuronal markers FoxG1 and Map2. One hundred percent of iNs were stained for FoxG1 and Map2 on d 4. Scale bar, 100 μm. E) Heatmap of gene expression levels of cortical layer markers and neuronal markers in iNs at different days after induction. All expression levels are normalized to those on d 5 (D5). EF1α, elongation factor-1 alpha; PuroR, puromycin resistance gene; rtTA3, reverse tetracycline-controlled transactivator 3; TRE, tetracycline response element.