FIGURE 7.

Human U-87 MG cells expresses CMKLR1 and CCRL2, and chem157S causes calcium signaling. A, mRNA levels of chemerin and its receptors, CMKLR1, CCRL2, and GPR1 in human glioma cell lines 1321N1, DBTRG-05MG, Hs 683, T98G, U-87 MG, and Jurkat cells were quantified with SYBR Green-based real-time quantitative PCR. The data are expressed as mean ± S.D. of relative gene expression to the Hs 683 cell line. B, U-87 MG cells were analyzed for the expression of human CMKLR1 and CCRL2 by flow cytometry. Isotype-labeled control is shown in purple, and anti-CMKLR1 or anti-CCRL2 shown in green. C, Fluo-8-loaded U-87 MG cells were stimulated with the indicated concentrations of chem157S (100 nm, ■; 30 nm, □; 10 nm, ▴; 3 nm, △; 1 nm, ●; and control, ○) and the fluorescent intensity monitored. The inset shows the maximum fluorescence intensity observed plotted as a function of the chem157S concentration. The data shown are representative of at least three independent experiments. The EC₅₀ was calculated based on the maximal fluorescence intensities observed at each concentration after stimulation.