Figure 1. Identification of ZnT3 oligomeric states.

A) Triton soluble extracts of PC12 cells (450 µg) co-expressing ZnT3-HA and ZnT3-myc, treated with and without cross-linker (DSP), were immunoprecipitated with HA (lanes 1 and 2), myc (lanes 4 and 5) or synaptophysin (Sphysin, lanes 7 and 8) antibodies. Western blots were probed with myc, HA and synaptophysin and ZnT3 antibodies, respectively. Control immunoprecipitation with synaptophysin antibodies fail to isolate ZnT3. Input 10 µg. B) 1.5 mg of Triton-X100 soluble supernatant of ZnT3-myc expressing cells treated in the presence of vehicle (DMSO) or DSP (+DSP) were separated by sucrose sedimentation. Fractions were collected from the bottom and analyzed by immunoblot with myc antibodies. An 80 kDa and high molecular weight forms of ZnT3 were observed, together with the monomeric 40 kDa species.