Fig. 1. Baculoviral constructs of MAP2 expressed in Sf9 cells. (A) The three open boxes represent the three repeat sequences involved in the microtubule-binding activity of MAP2. The highlighted parts of the sequence correspond to the highly conserved regions. The percentage of cells with processes is indicated for each MAP2 construct. For details on the morphological analysis see Materials and Methods. (B) Sf9 cells were infected with the recombinant baculovirus containing the truncated forms of MAP2 at an m.o.i. of 5.0. At 72 hours post-infection, the cells were lysed and 30 µg of cell lysates was separated on SDS-PAGE (7.5%). The proteins were transferred to a nitrocellulose membrane and the expression of the constructs was revealed using the anti-MAP2 antibody HM2, except for the construct corresponding to the microtubule-binding domain (Mt), which was revealed with the antibody 46.1. The molecular mass of the standards are indicated on the left: myosin (209 kDa), ß-galactosidase (134 kDa), bovine serum albumin (84 kDa), carbonic anhydrase (40 kDa), soybean trypsin inhibitor (32 kDa) and lysozyme (19 kDa). In lane MAP2b, MAP2b-1 and Prob, the multiple bands are caused by the protein degradation that occasionally occurs during preparation of cell lysates.