FIG. 1.

Characteristics of BM-derived EPCs and localization of TRPC1 in primary EPCs. (A) Representative BM-derived EPCs exhibited a cord-like (A1), tubular-like (A2), or cluster-like (A3) appearance (arrows). (B) EPCs showing uptake of acetylated LDL (red) and lectin (green) binding (90.7%±1.2%, n=3; 3 random fields per well).(C) Flow cytometry analysis of primary EPCs cultured for 4–7 days. Cells labeled with fluorescent antibodies recognizing CD133, VEGFR-2, CD34, and CD45 are shown as light green areas. The corresponding negative controls are exhibited as the white area on each box, the lines represent a positive gate, and numbers indicate the percentage of positive cells. (D) Subcellular localization of TRPC1 in primary EPCs. Top: Primary EPCs were incubated with antiTRPC1 polyclonal antibodies and Cy3-conjugated secondary antibodies. Analysis of the red fluorescent signal (TRPC1) indicates that TRPC1 is predominantly localized to the plasma membrane, with minor staining in the cytoplasm of EPCs. The scale bar represents 25 μm. Bottom: Control EPCs without primary antibody displayed no red fluorescent signal. BM, bone marrow; DAPI, 4, 6-diamidino-2-phenylindole; EPCs, endothelial progenitor cells; TRPC1, transient receptor potential canonical-1; LDL, low-density lipoprotein.