Fig 6.

Generation of P. falciparum lines expressing intact or disrupted MSPDBL2 in a majority of schizonts. (A) Schematic representation of the primary structural characteristics of MSPDBL2, as well as the TdTOM-3×HA tagged replacement (MSPDBL2-TAG) and the 3×HA-tagged disrupted version (MSPDBL2 DEL), both at the endogenous locus in the different SLI lines. The N-terminal signal sequence, the DBL, and SPAM (“secreted polymorphic antigen associated with merozoites”) domains and the location of the recombinant protein sequences used to raise the α-N-terminal and α-C-terminal antibodies are indicated. G418 treatment was initially used to select the neomycin-resistant engineered parasites, and cultures were treated with a fresh round of G418 for at least 6 days immediately prior to phenotyping assays. (B) As intended, most schizonts expressed either the entire or truncated version of MSPDBL2 in the respective lines, as assessed by immunofluorescence using antibodies against N-terminal or C-terminal MSPDBL2 sequences. All individual counts and assay values are given in Table S8. (C) Representative immunofluorescence (red) microscopy of schizonts expressing MSPDBL2 as an intact tagged version (MSPDBL2-TAG) or a disrupted version (MSPDBL2 DEL) stained by antibodies in murine sera against N-terminal (Nt) or C-terminal MSPDBL2 peptides (Ct). Parasite nuclei are stained with 4′,6-diamidino-2-phenylindole (DAPI), visualised as blue.