Fig. 3. Co-immunoprecipitation using α-Flag antibody and specific primary antibodies directed against TARDBP, FUS/TLS, NcoA3, NF45, cdc2 p34, and spectrin using total cell lysates, pre-fractionated nuclear fraction, or pre-fractioned cytoplasmic fractions of cells with or without the inducer. (A) Nuclear-Extract Kit was used to prepare nuclear, cytoplasmic, and total-cell extracts from cultured HEK293T/Flag-CTDSPL2 cells with or without inducer. (B) Co-immunoprecipitation using α-Flag antibody with total cell lysates, nuclear, or cytoplasmic fractions. Each extract was incubated with α-Flag-Protein A Agarose, washed, and subjected to 6-15% gradient SDS-PAGE. Gels were then immunoblotted with specific antibody. (C) Co-immunoprecipitation using specific antibodies against TARDBP, FUS/TLS, NcoA3, NF45, cdc2 p34, and spectrin. Spectrin is used as a false-negative control for immunoprecipitation.
