Journal: Nature Communications

Article Title: A basal body microtubule singlet-to-doublet transition in Plasmodium male gametogenesis

doi: 10.1038/s41467-025-64158-8

Figure Lengend Snippet: A U-ExM detection of the B tubule formation of MT doublet in the basal body and axoneme of gametocytes at 2 mpa. Parasites were co-stained with antibodies against PolyE and α/β-Tubulin. PolyE marks the mature B-tubule of MT doublet. White asterisk refers to the nuclear spindle. Scale bars = 5 μm. A representative from three independent experiments. The expansion factor is 4 ± 0.3. B Quantification of male gametocytes with PolyE labeled axonemal MTs in ( A ). Mean ± SD from three independent experiments, two-tailed t -test. C Diameter of axoneme in ( A ). n is the number of axonemes measured from three independent experiments. Mean ± SD, two-tailed t -test. D iU-ExM detection of the B tubule formation of MT doublet in the axoneme of gametocytes at 8 mpa. Parasites were co-stained with antibodies against PolyE and α/β-Tubulin. Scale bars = 5 μm. Insets show enlargements of the boxed areas with a cross-section view of the axoneme. Scale bars = 1 μm. A representative from three independent experiments. E TEM of axonemal MT architecture in the gametocytes at 8 mpa. Scale bars = 1 μm. Insets show enlargements of the boxed areas with a cross-section view of the axoneme. Scale bars = 100 nm. A representative from three independent experiments. F Quantification of male gametocytes with normal “9 + 2” MT structure axoneme. Mean ± SD from three independent experiments, two-tailed t -test. G Percentage of three types of MT structure in ( E ). Intact MT doublet (A + B tubule), incomplete MT doublet (A tubule + incomplete B tubule), and MT singlet (A-tubule or Central pair (CP) MT singlet). Mean ± SD from three independent experiments. H Cartoon showing the δ-Tub or ε-Tub deficiency in the B-tubule formation of MT doublet in the basal body. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies include: rabbit anti-HA (Cell Signaling Technology, cat#3724S, 1:1000 for immunoblotting IB, 1:1000 for immunofluorescence IF), mouse anti-Myc (Cell Signaling Technology, cat#2276S, 1:1000 for IF, 1:1000 for IB), mouse antibody against both α-Tubulin I and α-Tubulin II (Sigma-Aldrich, cat#T6199, 1:1000 for IF), mouse anti-β-Tubulin (Sigma-Aldrich, cat#T5201, 1:1000 for IF), rabbit anti-Polyglutamate chain (PolyE) (AdipoGen, cat#AG-25B-0030, 1:1000 for IF).

Techniques: Staining, Labeling, Two Tailed Test