Fig. 1. Bacterial overexpression and purification of citrin, aralar1 and their C-terminal domains. Proteins were separated by SDS–PAGE and either stained with Coomassie Blue dye (A and C) or transferred to nitrocellulose membrane and detected with specific polyclonal antibodies (B, D and E). (A and B) Cells of E.coli CO214(DE3) overexpressing citrin. Samples were taken at induction of expression (lane 1) and 12 h later (lane 2). The same number of bacteria was analyzed in each sample. Lanes 3, 6 µg (A) and 1 µg (B) of purified citrin obtained from bacteria in lane 2 of (A). (C) A 5 µg aliquot of purified citrin (lane 1), its C-terminal domain (lane 2), aralar1 (lane 3) and its C-terminal domain (lane 4). (D) Immunoreaction of 0.2 µg of purified citrin and its C-terminal domain (lanes 1 and 2, respectively) with antiserum to the C-terminal domain. (E) Immunoreaction of 0.2 µg of purified aralar1 and its C-terminal domain (lanes 1 and 2, respectively) with antiserum against the C-terminal domain.