Figure 4.

Patterns of glycosylation of cell-associated and secreted LHCGR recombinant proteins. a), the relative position of the glycosylation sites in the three recombinants; b), the LHCGR 229-318 residue proteins from transfected cell extracts (lane 1-9) as well as concentrated culture supernatants (lanes 10-18) at 96h following transfection were digested with Endo-H and PNGaseF prior to Western blot analysis as described; notably the protein isoforms resistant to deglycosylation by both enzymes were not secreted; c), both unglycosylated and glycosylated LHCGR 291 and LHCGR 318 migrate faster under non-reducing (UR) compared to reducing (R) conditions suggesting that folding of the proteins is independent of glycosylation. Lanes 1-3 are longer exposure of lanes 4-6.