Fig. 3. Characterization of unclassified cells (UCs).

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Fig. 3. Characterization of unclassified cells (UCs). Characterization of unclassified cells (UCs). (A) Analysis of the mesenchymal markers CD29, CD44, CD51, CD73, CD90.2, CD105, CD106, CD140a and CD166 among CD45−Ter119−CD31−LYVE-1− unclassified cells (n = 3). (B) Spleen sections from 16.5 dpc were stained with CD51 (green) and TOTO-3 (blue). Scale bar: 20 µm. (C) A single cell suspension was obtained from fetal spleen at 16.5 dpc. CD45−Ter119−CD31−LYVE-1−CD51+ defines CD51+ cells. Relative expression (RQ) of Scf and Igf1 mRNAs was examined by real-time PCR in CD51+ cells (n = 3). Hematopoietic cells (HCs) served as reference controls. (D) Amounts of SCF and IGF-1 protein per 100,000 cells in CD51+ cells. Expression of SCF and IGF-1 was higher in CD51+ cells than in HCs (n = 3). (E,F) Immunohistochemistry of endothelial and mesenchymal cell markers, CD31 and CD51, respectively stained with SCF and IGF-1 at 16.5 dpc. Samples were observed under confocal microscopy. CD31+/− and CD51+/− cells expressed both SCF and IGF-1. Scale bars: 20 µm. Data are mean±standard deviation (SD). *P<0.05. Keai Sinn Tan et al. Biology Open 2015;bio.201410686 © 2015. Published by The Company of Biologists Ltd