Gametocytes Immunofluorescence Assay

Gametocytes immunofluorescence assays were performed as previously described (Volkmann et al., 2012 (link)). For HA, c-myc and α-tubulin staining, purified cells were fixed with 4% paraformaldehyde and 0.05% glutaraldehyde in PBS for one hour, permeabilised with 0.1% Triton X-100/PBS for 10 min and blocked with 2% BSA/PBS for 2 hr. Primary antibodies were diluted in blocking solution (rat anti-HA clone 3F10, 1:1000; polyclonal rabbit anti-c-myc ref C3956, 1:5000; mouse anti-α-tubulin clone DM1A, 1:1000, all from Sigma-Aldrich). Anti-rat Alexa594, anti-mouse Alexa488, anti-rabbit Alexa 488, Anti-rabbit Alexa594 were used as secondary antibodies together with DAPI (all from Life technologies, Switzerland), all diluted 1:1000 in blocking solution. Confocal images were acquired with a LSM700 or a LSM800 scanning confocal microscope (Zeiss).