TNFSF15-facilitated sFlt1 mRNA up-regulation and activation of PKC, Src, and Erk1/2 in EPC. (A) Changes of sFlt1 mRNA levels in response to TNFSF15 treatment in the presence or absence of 4-3H, GF109203X, or PMA, determined by real-time PCR. (B) Changes of sFlt1 mRNA levels in response to TNFSF15 treatment in the presence or absence of PP2, U0126, or LY294002, determined by RT-PCR. (C) Flow-cytometric analysis of Src phosphorylation in response to TNFSF15 treatment in the presence or absence of 4-3H, GF109203X, or U0126. Black lines, vehicle treated. Red lines, TNFSF15 treated. (D) Quantitative analysis of the data in C. Black bars, vehicle treated. White bars, TNFSF15 treated. (E) Flow-cytometric analysis of Erk1/2 phosphorylation in response to TNFSF15 treatment in the presence or absence of 4-3H, GF109203X, or PP2. Black lines, vehicle treated. Red lines, TNFSF15 treated. (F) Quantitative analysis of the data in E. Black bars, vehicle treated. White bars, TNFSF15 treated. (G) Microscopic images of Jmjd6 immunofluorescent staining of cells treated with TNFSF15 or vehicle for 6 h in the presence or absence of 4-3H; red, Jmjd6; blue, DAPI-stained nuclei. (Scale bar, 20 μm.) (H) Fluorescence intensity of Jmjd6 in TNFSF15-treated cells in the presence or absence of 4-3H. (I) Changes of Jmjd6 fluorescent intensity in EPC in response to TNFSF15 treatment. The experiments were repeated two times. Data are mean ± SD. **P < 0.01, Student t test.