Figure S1. Immunoblot of PrPC antibodies in mouse and guinea pig tissues. PrPC expression in brain and ileal homogenates (10% w/v) were prepared and visualized as previously described.40 (A) Immunoblotting, using SAF-32 or SAF-83 antibodies, was used to determine PrPC expression in whole brain homogenates of Prnp+/+ mice (M) or guinea pigs (GP). PrPC expression levels, as determined by SAF-83 or SAF-32, were comparable in mice. In contrast, PrPC expression as determined by SAF-32 was slightly reduced, whereas SAF-83 was absent, in the guinea pig. (B) PrPC expression in Prnp+/+ mice or guinea pigs was determined in 3 cm full-thickness segment homogenates of the terminal ileum. The expression of PrPC using these two antibodies mirrored that as described for the brain. Note the variability at potential N-glycosylation sites between the brain and the ileum. β-actin was used as the lane loading control for all immunoblots.

Figure S2. Ileal PrPC expression in the submucosal plexus. (A) PrPC colocalized with PGP9.5 in ileal whole mount preparations from the Prnp+/+ mice (d-f, arrows). The most abundant colocalization was found to be on the cell surface (arrows) or on neuronal processes. PrPC immunoreactivity was absent in tissues of Prnp-/- mice (a-c). (B) Colocalization of PrPC and GFAP was limited in Prnp+/+ mice (d-f, arrows); PrPC was not detected in submucosal plexus preparations obtained from the Prnp-/- mice (a-c). PrPC antibody: SAF-83. n ≥ 12; Scale bar: 25 μm.

Figure S3. Ileal expression of PrPC and synaptophysin in the submucosal plexus of mice and guinea pigs. (A) PrPC colocalized with Synp in the ileal submucosal plexus of Prnp+/+ mice (d-f, arrows), but was absent in Prnp-/- mice (a-c, n = 6). (B) PrPC expression in the submucosal plexus of the guinea pig (n =3). PrPC expressed within the submucosal ganglia colocalized with PGP9.5 (a-c), GFAP (d-f) and Synp (g-i). PrPC antibody: SAF-83. Scale bar: 25 μm.

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