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. 2023 Jan 25;26(2):106056. doi: 10.1016/j.isci.2023.106056

Figure 3.

Figure 3

B9 localizes to a subset of sporozoite micronemes and is secreted upon parasite activation

(A) Immunofluorescence analysis of PbGFP and PbB9-Flag sporozoites labeled with anti-Flag antibodies (red). Parasites express GFP (direct detection, green) and nuclei were stained with Hoechst 33342 (blue). Scale bar, 10 μm.

(B) Localization of B9 in sporozoites. First panel, confocal image of GFP (direct detection, green); second panel, visualization of B9-Flag (with anti-Flag, red) using 2D STED (maximum intensity projection). Scale bar, 2 μm.

(C) STED images of sporozoites expressing B9-Flag and P36-mCherry, labeled with anti-mCherry (red) and anti-Flag (cyan) antibodies. Scale bars, 2 μm (200 nm in insets). Some micronemes are labeled for both B9 and P36 (white arrowheads). Others are stained only for B9 (cyan arrowheads) or P36 (red arrowheads).

(D) Immunoblot of B9-Flag sporozoite pellets and supernatants in control conditions (4°C) or after stimulation of microneme secretion (37°C), using anti-Flag or anti-GFP antibodies. The data shown are representative of three independent experiments. See also Figure S5.