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. 2013 Aug 5;110(34):13863–13868. doi: 10.1073/pnas.1304529110

Fig. 3.

Fig. 3.

TNFSF15-facilitated down-regulation of mFlt1 and up-regulation of sFlt1 in EPC cultures. (A) Flow cytometry histogram of Lin−-Sca-1+ EPC treated with TNFSF15 (bold line) or vehicle (thin line). The cells were labeled with a phycoerythrin-conjugated anti-mFlt1 antibody or an isotype-matched antibody (dotted line). (B) Percentages of mFlt1+ cells in TNFSF15-treated cultures in the presence or absence of the TNFSF15 neutralizing antibody 4-3H, determined by flow cytometry. (C) Percentages of mFlt1+ cells in TNFSF15 (squares)- or vehicle (circles)-treated cultures in the presence of 4-3H at indicated concentrations. (D) Western blotting analysis of mFlt1 and sFlt1 protein levels in TNFSF15- or vehicle-treated cultures at the indicated time points. (E) Concentrations of sFlt1 in culture media determined by ELISA at indicated time intervals following TNFSF15 treatment in the presence or absence of 4-3H; black, vehicle; crosses, 4-3H alone; white, TNFSF15 alone; slashes, TNFSF15 and 4-3H. (F) mRNA levels of mFlt1 and sFlt1 in TNFSF15- or vehicle-treated cultures at the indicated time points, determined by RT-PCR; black, mFlt1 in vehicle-treated cells; crosses, mFlt1 in TNFSF15-treated cells; white, sFlt1 in vehicle-treated cells; slashes, sFlt1 in TNFSF15-treated cells. The experiments were repeated two times. Data are mean ± SD. *P < 0.05, **P < 0.01, Student t test.