Fig. 5
Infection of P2FJ6 cells with ovine MM2-SCJD prions, and PrPres glycopattern of the cell-passaged prions in tg338 mice. (a-c) Western blot analysis of PrPres in P2FJ6 cell lysates infected with brain (Br) and spleen (Sp) extracts of tg338 mice infected with MM2-sCJD prions, either uncloned (MM2) or cloned (Cl.MM2), and mice challenged with either brain or spleen extracts of PMCA MM2-sCJD prions (MM2 → PMCA → tg338). The results are shown over 3 (a) or 4 passages (b, c) post-cell exposure, except in lanes 11-12 in (A), where the analysis was made at 14 passages. The gels in b and c are overexposed purposely to reveal accumulation of T1Ov PrPres after infection with spleen material. Untransfected RK13 cells (RK13, arrow in a) did not accumulate detectable levels of PrPres. Infection with 127S prions was performed in parallel to estimate the efficacy of infection. The gels were loaded with 250 μg of PK-digested protein or 80 μg for 127S infection. The blots were probed with Sha31 antibody. (d) Western blot analysis of PrPres in the brain and spleen tissue of tg338 mice challenged with P2FJ6 cell lysates infected with uncloned (MM2), cloned (Cl.MM2) MM2-sCJD prions, PMCA MM2-sCJD prions (MM2 → PMCA → tg338), and 127S as control. The equivalent of 1 and 2 mg of brain and spleen tissue was loaded on the SDS-PAGE gels. The blots were probed with Sha31 antibody