FIGURE 5.

Western blots of PfACP from P. falciparum cell culture under normal and oxidizing conditions. (1) Reduced PfACP runs just under 10 kDa, and PfACP with its apicoplast transit peptide can be seen as a minor band at 15 kDa. (2) Parasites in culture medium were treated with H₂O₂ at 0 mM, (3) 10 mM, and (4) 100 mM. The large low molecular weight band is residual heme reacting directly with the ECL substrate. (5) Parasites liberated from their red blood cells were treated with H₂O₂ at 10 mM and (6) 20 mM. (7) Air oxidized recombinant PfACP provides a marker for monomer, dimer, and trimer. (8) P. falciparum parasites were treated with 20 mM diamide for 30 min at 4 °C, then lysed without reducing agent or iodoacetamide, and analyzed by Western. (9) The same sample used in lane 8, except 300 mM DTT was added before electrophoresis. Lanes 6 and 7 are from the same blot, and lanes 8 and 9 are from the same blot. Exposure settings are similar across gels, and identical within them.