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. 2018 Nov 27;8:413. doi: 10.3389/fcimb.2018.00413

Figure 3.

Figure 3

P36 and P52 are Localized in the Sporozoite Micronemes. (A) Representative immunofluorescence microscopy images of P36mCherry salivary gland sporozoites showing the internal localization of P36mCherry compared to the micronemal proteins P52 and TRAP and in contrast to the inner membrane complex protein MTIP. Second row shows signal co-localization with α-mCherry and α-PyP36 respectively. Nuclei were stained with DAPI. Primary antibodies used: rat α-mCherry, rabbit α-MTIP, mouse α-PyP36.1, mouse α-PyP52 (13G10), mouse α-PyTRAP.2. Secondary fluorescently-labeled antibodies: α-rat-AF594, α-rabbit-AF488, and α-mouse-AF488 (Scale bar: 5 um). (B) Representative electron microscopy images of P36mCherry sporozoites show the co-localization of P36, P52 and TRAP in the micronemal organelles. Top left: P36mCherry (α-PyP36.1) and P52 (α-P52-2D5) are shown in the micronemes of individual sporozoites (15 nm gold). Top right: dual labeling with rabbit α-PyP36 (S) and mouse α-PyP52 (2D5) (L). Bottom left: dual labeling with rabbit α-PyP36 (S) and mouse α-PyTRAP.2 (L). Bottom right: dual labeling with rabbit α-TRAP (S) and mouse α-PyP52 (2D5) (L). The chart shows the percentage of micronemes with single and dual labeling in the analyzed sporozoites (Figure S6). (S) = 10-nm gold; (L) = 15 nm gold (Scale bar: 500 nm).