Figure 2

Stably transfected breast cell lines can be generated with functional GnRH receptor. Relative levels of GnRH at the cell surface detected by ligand binding assay in human cell lines stably transfected with rat GnRH receptor cDNA expression construct A. Subclones of MCF-7 clone 30 expressing modified levels of GnRH receptor at the cell surface were isolated. HEK293 and T47-D10 cells demonstrated background levels of binding. All other cell lines shown demonstrated significantly (p < 0.05 ANOVA) higher levels of specific binding. B. MCF-7-30-7 was subcloned from MCF-7-30 and then transfected with a PvuII SV40-hygromycin resistance gene fragment. Clones resistant to G418 and hygromycin were screened for altered GnRH receptor expression. GnRH receptor levels were elevated in clone MCF-7-30-hygro14, similar to levels in HEK293_[SCL60]. * p < 0.05 (ANOVA followed by Dunnett's test) indicates significantly higher in MCF-7-30-7 and MCF-7-30-hygro14 relative to MCF-7-30 binding.