Infectious Prions Accumulate to High Levels in Non Proliferative C2C12 Myotubes
Figure 2
(A) Time course of PrPSc accumulation. Confluent myotubes were infected by incubation of cells in media spiked with 100 µL of 1% RML brain homogenate or mock infected with normal brain homogenate overnight. Media was changed daily. After 5, 10 or 15 days in culture, cells were lysed and analyzed for PK-resistant PrP. (B) Myoblasts were infected by incubating in media spiked with 100 µL of 1% RML brain homogenate or normal brain homogenate. Cells were analyzed after passages 7, 8 and 9 which correspond to myotube days 11, 13 and 15 respectively and no PrPSc was observed. For both panels (A,B), 7.5 µg of protein was loaded in PK- lanes. 15 µg of protein equivalent was loaded in lanes treated with PK. 10 µl of 0.15% BH was loaded ∼11.25 µg protein. Anti-PrP antibody SAF 83 was used.