(a) A representative example of a schizont from a clinical P. falciparum isolate with PfRh4 expression, as determined by immunofluorescence microscopy. Order of panels: phase, DAPI stain (labels nucleus), anti-Ron4-Alexa 488, anti-Rh4-Alexa-597, overlay. (b) Western blots showing expression of PfRh4 by P. falciparum isolates recently adapted to in vitro culture. Proteins were extracted from schizonts and the membrane was probed with anti-PfRh4 antibodies (upper panel) and anti-HSP70 antibodies (lower panel) as a loading control. The positions of molecular weight standards are indicated on the left. Isolates tested were 3D7, XIE (from PNG), Pf2004 (Ghana), Pf2006 (Ghana), HCS3 (Thailand) and a W2mef-PfRh4 knockout line (Rh4ko).