Figure 5
Natural Lepp12 protein is non-secreted protein located in promastigote nucleus. Western blotting was performed using immunopurified anti-Lepp12 antibodies (at 2 microg/ml) on total promastigote 3 % SDS lysate (positive control, lane 1), 20 % NP40 lysate (lane 2), cytosol (s1, lane 3), cell membranes (c1, lane 4), nuclear extract prepared as described in Methods section before (lane 5) or after (lane 6) 3% SDS lysis, and PEG concentrated culture supernatant (lane 7). Proteins were prepared from 1.6 × 108 cells. In each lane, 30 microg of proteins were separated on a 14% polyacrylamide SDS-gel, corresponding to an equivalent of 4 millions (lane 1), 8.6 millions (lane 2), 5.3 millions (lane 3), 4.8 millions (lane 4), 18.5 millions (lane 5), 12 millions (lane 6) and 2.2 millions (lane 7) promastigotes. Molecular mass markers are indicated in kDa.