Fig. 6: GnRH inhibits multiple beneficial functions of stem cells in vivo.

Schematic representation of the experimental protocol as described in the materials and methods section (a). Mice were treated daily for 21 days with GnRH (0.1 mg/kg, intraperitoneally) or vehicle (PBS). Stem cells were isolated from mouse adipose tissue, and changes in cell viability were determined by an MTT assay. Stem cell viability (%) was calculated as a percent of the vehicle control (b). The changes in migratory capacity were measured via the transwell assay (c) and western blotting for MMP-2 and MMP-9 (d). The changes in osteoblast differentiation were determined by alizarin red staining. The relative quantification of calcium mineral content was performed by measuring the absorbance at 570 nm (e). Real-time PCR results showed the changes in the expression of the mouse stem cell markers C-MYC and KLF4 after GnRH treatment in vivo (f). β-actin was used as the internal control. The data are presented as the mean ± SD of three independent experiments