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. 2022 Dec 28;2022:1396735. doi: 10.1155/2022/1396735

Figure 2.

Figure 2

Crispr-Cas9-mediated targeted integration of GFP at the AAVS1 site. (a) Schematic diagram of the knock-in strategy showing the GFP-containing homologous recombination sequence (hAAVS1-CMV-GFP) and the insertion site (hAAVS1 locus). (b) Flow cytometric analysis results of iNSC-GFP and control iNSCs without GFP knock-in. (c) Immunofluorescence staining for GFP, NESTIN, KI67, PAX6, and SOX2. Scale bars, 75 μm. (d) The percentages of iNSCs at passage no. 40 with or without GFP knock-in that were positive for SOX2, KI67, NESTIN, PAX6, and GFP. (e) Expression of SOX1, SOX2, PAX6, HES5, FABP7, and β-actin of 293T, iNSCs of passage no. 20, and iNSC-GFP of passage nos. 20, 25, 30, and 40 at the transcription level.