FIG. 1.

Coimmunoprecipitation of LC8 and P protein of Mokola virus. Pmok and LC8 genes were transcribed and translated in vitro using the TnT coupled reticulocyte lysate system (Promega) in the presence of [³⁵S]methionine (>1,000 Ci/mmol; Amersham). The translated products (A, lane 3) were subjected to immunoprecipitation with protein A-Sepharose beads loaded with polyclonal anti-LC8 antibodies (A, lane 2) or with anti-Flag monoclonal antibody M2 (B, lane 2) or without any antibody (A, control lane 4, and B, control lane 3). Lanes 1, molecular size marker. Immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (20% polyacrylamide). (C) Ex vivo expression in Neuro 2A cells transfected with tagged LC8. The cells were labeled with [³⁵S]methionine, and cell extracts were immunoprecipitated with anti-Flag monoclonal antibody M2 (lane 3). A similar experiment was performed after cotransfection with the Pmok WT (lane 2). Immunoprecipitation of cell extracts from Neuro 2A cells cotransfected with the LC8 WT and tagged Pmok plasmids, infected with Mokola virus (multiplicity of infection, 10) (lane 4).