Journal: bioRxiv

Article Title: A divergent cyclic nucleotide binding protein promotes Plasmodium ookinete infection of the mosquito

doi: 10.1101/2025.01.30.635676

Figure Lengend Snippet: PbCBP-O is required for efficient midgut traversal. (A) PbΔCBP-O ookinetes show regular motility. Average speed is 3.07 and 3.2 μm/min for wild-type and ΔCBP-O, respectively (wild-type n = 38; PbΔCBP-O n = 48, unpaired t-test). (B) Comparable ookinete maturation in the mosquito midgut is observed in PbΔCBP-O and wildtype parasites. Nineteen or twenty-one hours post-infection, six midguts were scored per replicate (n = 2). Significance was analysed using an unpaired t-test. (C) Expression of micronemal proteins including CTRP, WARP and chitinase are unaffected in the absence of CBP-O. Actin is used as loading control (left panel). Chitinase secretion is maintained in PbΔCBP-O ookinetes. Western blot of culture supernatants from wild type and PbΔCBP-O ookinetes show similar levels of chitinase secretion (right panel). (D) Immunofluorescence showing decreased ookinete presence in the basal side of mosquito midguts. Midguts were fixed and stained with P25 (plasma membrane marker) and DAPI 24 hours post-infection. P25 + parasites were quantified, and single dots show number of ookinetes counted per midgut (n = 20 midguts per genotype, unpaired t-test.). Scale bar = 200μm. In all relevant panels, statistical significance is indicated as: ns = not significant and **** = p < 0.0001.

Article Snippet: The slides were mounted using 30μl of Fluoromount-G with DAPI (Fisher Scientific) and a 40mm coverslip and left overnight to set.

Techniques: Infection, Expressing, Control, Western Blot, Immunofluorescence, Staining, Membrane, Marker