The PfRH protein family of Plasmodium falciparum is implicated in erythrocyte invasion. Here we report variations in the sequence, transcription, and protein expression of four different members of this family in three parasite lines, 3D7, T996, and FCB1. There are sequence polymorphisms in PfRH1, PfRH2a, PfRH2b, and PfRH3, ranging from variations across repeat regions to a 585-bp deletion in the 3' end of PfRH2b in T996. Not all the genes are transcribed: although all members of the family are transcribed in 3D7 and T996, PfRH2a and PfRH2b are not transcribed in FCB1. The PfRH1, PfRH2a, and PfRH2b proteins are expressed in late schizonts and merozoites and are located in apical organelles and on the apical surface. However, the PfRH1 protein does not appear to be correctly targeted to the apex in 3D7 and T996. In contrast, the PfRH1 protein is present at the apical end of FCB1 merozoites, but the PfRH2a and PfRH2b proteins are undetectable. The apparent redundancy in the PfRH family of proteins at the level of gene number and sequence and the variations in transcription and protein expression may allow the parasite to use alternative invasion pathways.