Fig. 3

PfPM VII and PfPM X expression in Plasmodium falciparum sexual stage parasites demonstrated by IFA and western immunoblot. IFA of in vitro-cultivated parasites was performted using primary antibodies 1B4, 2B1, 6A9, 3G6 or isotype IgG2b and subsequently labelled with Alexa Fluor 488-labelled anti-mouse antibodies (green); nuclei are visualized with DAPI (blue). a IFA of in vitro-cultivated P. falciparum demonstrated PfPM VII expression in zygotes (Z) and ookinetes (O). b IFA of in vitro-cultivated P. falciparum demonstrated PfPM X expression in zygotes (Z) and ookinetes (O) but not gametocytes (G). c Western immunoblot analysis of total protein isolated from P. falciparum sexual stage parasites. Antibodies directed against PfPM VII recognized a ~46 kDa protein expressed in ookinetes (O) but not gametocytes (G). This protein is between the predicted sizes of full length PfPM VII at 52 kDa and the catalytic domain at 43 kDa. d Western immunoblot analysis of total protein isolated from P. falciparum sexual stage parasites. Both 6A9 and 3G6 recognized protein expressed in mixed gametes and zygotes (m/Z) and ookinetes (O) but not gametocytes (G). Bands are consistent with the predicted sizes of full length PfPM X at 61 kDa (←F), the PfPM X catalytic domain at 36 kDa (←C), and the PfPM X proenzyme domain at 23 kDa (←P). IgG isotype control did not recognize the 61 kDa or 23 kDa bands on western immunoblot but continued to recognize the bands at 42–55 kDa and 27–30 kDa