FIG 3.

enRep-M9l encodes a protein with cytoplasmic localization. The expression of enRep-M9l isoforms was analyzed by Western blotting and immunofluorescence. (A) NIH 3T3 cells were transfected with plasmids encoding Flag-enRep-M9lL, Flag-enRep-M9lS, enRep-M9l-FlagL, enRep-M9l-FlagS, or Flag-Odegus4 or with empty vector. At 48 h later, cells were lysed, and Western blot assays were performed using anti-Flag or anti-tubulin antibodies as a loading control. The migration of the molecular marker is shown on the left-hand side, and the antibody used for each blot is indicated on the right-hand side. S, short; L, long. (B) NIH 3T3 cells seeded over coverslips were transfected with plasmids encoding Flag-enRep-M9lS, enRep-M9l-FlagS, Flag-Odegus4, or mouse GFP-Myosin9. At 24 h later, cells were fixed, and immunofluorescence was performed using anti-Flag or anti-GFP antibodies. DAPI was used to stain the nucleus. Bar, 10 μm.