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. 2009 Jun 5;8(8):1134–1145. doi: 10.1128/EC.00083-09

FIG. 5.

FIG. 5.

(A) Membrane extraction of 3D7PfsD1-1-infected erythrocytes. STris, supernatant after 50 mM Tris lysis; SCarb, SHS, and SUrea refer to supernatant after extraction with, respectively, carbonate, high salt, or urea buffer; P, final membrane pellet. In all cases PfsDer1-1-GFP can be detected exclusively in the final membrane pellet fraction, suggestive of a tight membrane association. Control proteins PfHsp70 (soluble protein) and PfExp1 (protein containing a TM domain) can be detected in the STris and P fractions, respectively, as expected. (B) Triton X-100 (T-X-100) extraction of 3D7PfsD1-1-infected erythrocytes. P, pellet after extraction; S, supernatant following extraction. PfsDer1-1-GFP can de detected in the pellet fraction only after treatment, whereas PfExp1 is solubilized already with 0.5% Triton X-100. (C) Developmental expression of PfsDer1-1-GFP. Protein extracts from highly synchronized 3D7PfsD1-1 parasites were prepared and analyzed by Western blotting with anti-GFP antibodies. As a control, we analyzed expression of PfHsp70 using anti-PfHsp70 antibodies. Expression of PfsDer1-1-GFP appears (in comparison to PfHsp70) to gradually increase during the 48-h intraerythrocytic life cycle of the parasite. Time scale refers to hours postinvasion. Troph, trophozoite; Schizon, schizont; α, anti.