Figure 1.

The effects of erlin1 and erlin2 deletion on assembly. A, lysates were made from control, E1KO, and E2KO αT3 cells and probed in immunoblots for ubiquitin, IP₃R1, p97, Hrd1, gp78, erlin1, erlin2, and RNF170. B, nondenaturing PAGE of cell lysates probed with anti-erlin1, anti-erlin2, or anti-p97 as a loading control. C, anti-erlin1 or anti-erlin2 immunoprecipitates (IP) from control, E1KO, and E2KO αT3 cells probed for the proteins indicated. D, membrane preparations from control, E1KO, and E2KO αT3 cells (lanes 1–3) and immunoprecipitates from control αT3 cells, incubated either without (lanes 4–6) or with 100 nm GnRH for 5 min (lane 7), were probed in immunoblots for the proteins indicated, including erlin1 and erlin2 with anti-erlin^pan (bottom panels). Note that several erlin-unrelated background bands were recognized by anti-erlin^pan even in the membrane preparations (lanes 1–3) but that these were not present in immunopurified material (lanes 4–7).