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Figure 1

Susceptibility to scrapie infection and PrP level of various sublines of N2a cells. N2a populations as propagated routinely in the lab and single clones transformed with a PrP expression or a control vector were seeded into 24-well plates (2 10⁴ cells per well) and grown to confluence. (a) Cultures were exposed for 3 days to purified mouse PrP^Sc (RML strain, 20 ng/ml), cultured for 29 days (eight passages), and assayed for PrP^Sc formation by the cell blot assay (14). (b) Prion-susceptible N2a/Bos2 and resistant N2a/2M11 cells were exposed for 3 days to the dilutions indicated of infected 10% brain homogenate, cultured for 14 days (three passages), and assayed for PrP^Sc formation. Cells exposed to a 10⁴ dilution are still slightly positive. (c) Western blot analysis of N2a sublines was performed with monoclonal anti-PrP antibody 6H4 as outlined in Materials and Methods. Cells transfected with the expression plasmid for mouse PrP^c, MHM2 PrP (21) or MH2M PrP (21) are indicated by mo, M2, or 2M, respectively. BOS designates cells cotransfected with pSVneo and pEF-BOS-EX. N2a, the original uncloned cells, as well as the highly susceptible N2a/Bos2 cell line show similar low expression of PrP^C as compared with the nonsusceptible mo5 or 2M11 lines.