Fig. 2. Generation of iPSCs from an AD patient harboring a PSEN1 (E120K) mutation, and an eldely normal subject. (A) Established iPSC lines from both control and PS1-E120K patient showing the expression of pluripotent stem cell markers, such as OCT4 (red), SOX2 (green), SSEA4 (red) and TRA-1-81 (red). (B) Reverse transcription PCR (RT-PCR) showing the expression of pluripotency markers (OCT4, SOX2, NANOG, SSEA4 AND TRA-1-81) in both iPSC lines. (C) Genomic DNA sequences showing the presence of the heterozygous E120K mutation (GAA to AAA) in the PSEN1 gene of the PS1-E120K-iPSC line. (D) Immunofluorescence analysis showing the potential of iPSC lines to form three germ layers, including ectoderm (type III β-tubulin [TUJ1], green), mesoderm (smooth muscle actin [SMA], green), and endoderm (α-fetoprotein [AFP], red). Scale bar: 100 µm. (E) Karyotype analysis of the control and PS1-E120K iPSC lines. (F) Reverse-transcription PCR analysis showing the absence of integration of the Sendai virus vectors. (G) PCR analysis showing no contamination by mycoplasma.
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