FIGURE 9.

Proliferation of stable cell groups in response to ghrelin. (A) Dose-dependent effects of ghrelin on stable SH-SY5Y-GHSR1a/OX1R cell proliferation. Cell proliferation was measured using a CCK-8 proliferation/viability assay. Stable cells that were not stimulated with ghrelin were used as the control. ^∗∗p < 0.01, cells co-expressing both GHSR1a/OX1R and treated ghrelin vs. control. (B) The stable SH-SY5Y-GHSR1a, SH-SY5Y-OX1R, and SH-SY5Y-GHSR1a/OX1R cells or SH-SY5Y cells were cultured in a 96-well plate and starved overnight in serum-free medium. Then, cells were stimulated with 100 nM ghrelin or 100 nM orexin-A or 100 nM ghrelin/orexin-A. Cell proliferation was measured as above. SH-SY5Y cells (Untransfected cells) were used as the control. The data points represent the mean ± SEM (N = 3) of four independent experiments. ^∗∗p < 0.01, cells co-expressing both GHSR1a/OX1R treated with 100 nM ghrelin vs. control.