The in vitro culture of the erythrocytic stages of P. falciparum was maintained as described by Trager and Jensen28 (link). Gametocyte production and enrichment was done as described by Fivelman et al.29 (link) and details are provided in Supplementary Methods . P. berghei ANKA parasites were maintained in BALB/c mice. PfISN1 gene was obtained by reverse transcription-PCR on parasite RNA.
The sequences of primers used for PCR amplification are provided in Supplementary Table5 . PfISN1 gene was cloned into pFCENv1 and pBCEN5 to obtain a C-terminal fusion with GFP. The resulting plasmids pFCENv1_PfISN1 and pBCEN5_ISN1GFP were used for transfection of P. falciparum and P. berghei, respectively using established protocols30 (link),31 (link) with details provided in Supplementary Methods . The confirmed lines of parasites carrying the PfISN1-GFP gene were examined by live-cell fluorescence microscopy using a Zeiss® LSM-510 META confocal microscope. Anti-PfISN1 antibody was raised in rabbits using purified recombinant PfISN1 and affinity purified using Sepharose beads conjugated to PfISN1. Using the antibody, indirect immunofluorescence microscopy was carried out using a Zeiss® LSM-510 META confocal microscope to check the localization of PfISN1 in intraerythrocytic stages of P. falciparum.
The sequences of primers used for PCR amplification are provided in Supplementary Table
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