Figure 8
Natural Lepp12 is present in promastigote nucleus as a phosphoprotein. Mixture of anti-phosphoserine and anti-phosphothreonine antibodies was used in the western blotting shown in order to evidence phosphoproteins. Enzymatic activity of anti-rabbit IgG peroxidase conjugate was revealed by chemiluminescence. Natural Lepp12 (nLepp12, lane 1) was immunoprecipitated from the promastigote nuclear extracts (corresponding to 3 × 109 cells) using anti-Lepp12 IgG (25 microg at 100 microg/ml). The immunoprecipitated material was Lepp12-specific as testified by control immunoprecipitation (cAb) performed with irrelevant IgG (lane 3). 15 microg of total promastigote lysate (proM, lane 2) was electrophorezed and electrotransfered onto nitrocellulose in parallel. Exposition time of MP-hyperfilm was 10 sec.