Supplemental Material

This article contains the following supporting material:

• Figure 1 - The LHRvariant with its endogenous signal peptide is not secreted from the HEK293 cells. Cells expressing the variant under the mouse metallothioneine-1 promoter were incubated in the absence or presence of 0.1 mM ZnCl₂ for 24 h to induce expression. Medium samples (lanes 3-4) and solubilized total cellular membranes (lanes 1-2) were subjected to hCG-affinity purification and aliquots of the eluates were analyzed by SDS-PAGE and Western blotting using a polyclonal antibody directed against the N-terminal domain of the rLHR. No specific variant forms were purified from the medium but two forms of M_r 52,000 and M_r 54,000 were detected in cells and their expression was increased about 3-fold in the presence of ZnCl₂.