Fig. 6: GCα knockdown in gametocytes results in gametogenesis defect.

a Diagram showing a promoter swap strategy to knockdown gcα expression in gametocytes, generating HA-tagged gcαkd mutant with endogenous gcα promoter replaced with the sera1 promoter. b Western blotting of GCα expression in asexual blood stages and gametocytes of the gcαkd parasite. The 6HA::gcα as a control. c Quantitative analysis of GCα protein expression in b. d Intracellular cGMP level in XA-stimulated gametocytes of the 17XNL and gcαkd parasites. Cells were incubated with 100 μM XA at 22 °C for 2 min before assay. Ctl are control groups without XA stimulation. e In vitro exflagellation rates for 17XNL, 6HA::gcα, and two clones of the gcαkd parasite after XA stimulation. f Day 7 midgut oocyst counts in mosquitos infected with 17XNL, 6HA::gcα, and two clones of the gcαkd parasites. Mosquito infection prevalence is shown above. g A proposed model of GEP1/GCα interaction essential for XA-stimulated cGMP synthesis and gametogenesis. Experiments were independently repeated three times in b, d, e, and f. Data are shown as mean ± SD in c, d, and e. Two-tailed unpaired Student’s t test in c, d, e, and f. Source data of c, d, e, and f are provided as a Source Data file.