FIG. 3.
Solubilization of GerD-FLAG from insoluble and P100 fractions. Spores of strain PP19 (gerD amyE::PsspB-gerD-flag) were disrupted, extracted, and fractionated, and fractions were treated with either 1% Triton X-100 or 0.5 M NaCl as described in Materials and Methods. Aliquots of fractions from equal amounts of spores were run by SDS-PAGE, proteins were transferred to a PVDF membrane, and GerD-FLAG was detected with anti-FLAG antibody. Samples run in the various lanes were as follows: 1, initial S100; 2, initial P100; 3, S100 from initial P100 plus Triton X-100; 4, P100 from initial P100 plus Triton X-100; 5, S100 from initial insoluble fraction plus Triton X-100; 6, P100 from initial insoluble fraction plus Triton X-100; 7, S100 from initial P100 plus NaCl; 8, P100 from initial P100 plus NaCl; 9, S100 from initial insoluble fraction plus NaCl; and 10, P100 from initial insoluble fraction plus NaCl. The numbered lines to the left of the figure denote the migration positions of molecular mass markers, in kDa, and the arrowheads to the right indicate the predicted migration positions of GerD-FLAG monomers (M), dimers (D), and trimers (T). Lanes 1 to 6 and 7 to 10 came from different gels.