Figure 2 | Scientific Reports

Figure 2

From: TALEN-mediated functional correction of human iPSC-derived macrophages in context of hereditary pulmonary alveolar proteinosis

Figure 2

Characterization of AAVS1.CSF2RAcoop iPSCs. (a) Immunohistochemical characterization of iPSCs. Healthy donor-derived hCD34iPSC16, PAP patient-derived iPSC20.1, as well as AAVS1.CSF2RAcoop clones #6 and #7 express typical pluripotency-related surface markers and transcription factors including SSEA4, SOX2, OCT4, Tra-1-60 and display alkaline phosphatase activity. (b) qRT-PCR-based gene expression analysis of pluripotency-related transcription factors relative to human GAPDH (housekeeping gene) and H9 ESCs used as reference. The transcription factors OCT4, SOX2 and NANOG are expressed by CD34iPSC16, the PAP20.1 as well as AAVS1.CSF2RAcoop #6 and #7. (H9, n = 3; CD34iPSC16, n = 3; PAP20.1, n = 3; #6, n = 2; #7, n = 2) (c) Gene expression analysis of CSF2RA coop. PAP20.1 was used as negative control iPSC line to verify CSF2RA coop expression in the edited AAVS1.CSF2RAcoop #6 and #7. (H9, n = 3; CD34iPSC16, n = 2; PAP20.1, n = 3; #6, n = 3; #7, n = 3) (d) Histograms depicting GM-CSF receptor (CD116) expression on iPSCs by flow cytometry. Mouse IgG1κ isotype controls (solid gray), CD116 (red); H9 ESC, H9 embryonic stem cells; DAPI, 4′,6-diamidino-2-phenylindole.

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