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. 2019 Jun 5;17:198–209. doi: 10.1016/j.omtn.2019.05.019

Figure 4.

Figure 4

FVIII Expression Was Restored in C-iPSC-Derived EPCs

(A) FACS analysis of the expression of the iEPC markers CD31 and CD34 on day 5 after differentiation. (B) Immunofluorescence staining of CD34 (red), CD31 (red), and CD144 (green) in C-iEPCs. Scale bar, 25 μm. (C) RT-PCR analysis of F8 expression in iEPCs. Primers based on exon 14 (E14) and primers based on exons 23 and 26 (E23–26) were used to detect F8 expression. GAPDH was used as a loading control. (D) FVIII antigen in iEPCs differentiated from hiPSCs was detected by ELISA. Bars represent the mean ± SEM (n = 3, independent cultures). (E) Immunostaining of endothelial markers CD31 and vWF in the mature iECs from iEPCs. (F and G) F8 expression in mature ECs derived from HA-iPSCs (HA-iECs), C-iPSCs (2-6-iECs and 2-46-iECs), and normal controls (N-iECs) was detected via immunostaining by an (F) N-terminal or (G) C-terminal FVIII antibody. DAPI was used for nuclear staining. FVIII-N, F8 protein, N-terminal. FVIII-C, F8 protein, C-terminal. Scale bar, 25 μm.