Figure 3.

GPI-anchored chimeric MHM2 PrP^C is released by PIPLC digestion but the transmembrane form is not. Chimeric PrP molecules denoted MHM2-Qa(−) for the transmembrane form and MHM2-Qa(+) for the GPI-anchored form were expressed in ScN2a cells. Media (lanes 1–6) and cell lysates (lanes 7–9) were collected from ScN2a cells stably transfected with MHM2-Qa(+) (lanes 1 and 2) or MHM2-Qa(−) (lanes 3, 4, and 8), or MHM2-FLAG-Qa (+) (lanes 5, 6, and 9). Cells were either digested with PIPLC (0.5 U) at 37°C for 4 h (even lanes), or without PIPLC (odd lanes). Western blot analysis was performed with α-PrP 3F4 mAb. Only the GPI-anchored MHM2-Prp was released into the media. Western blot analysis was performed with α-PrP 3F4 mAb.