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. 2021 Mar 15;11:634273. doi: 10.3389/fcimb.2021.634273

Figure 4.

Figure 4

PIMMS22 protein expression and localization. (A) Western blot analysis using α-GFP antibody on whole cell lysates of P. berghei c22::gfp parasites. The PbPIMMS22::GFP fusion protein band is indicated with a black arrowhead. The ANKA 2.34 parental parasite line was used as a negative control. Tubulin was used a loading control in all parasite stages except from midgut sporozoites where CSP was used. MBS, mixed blood stages; Gc(-), non-activated gametocytes; Gc(+), activated gametocytes; Ook, ookinetes; MgSpz, midgut sporozoites. Immunofluorescence assays of c22::gfp blood stage gametocytes treated or not treated with Triton X-100 (Tr.) (B), in vitro ookinetes (C), ookinetes traversing the mosquito midgut epithelium at 26 hpbf (D) and midgut sporozoites at 15 dpbf (E), stained with α-GFP (green), ookinete surface α-P28 (red), or sporozoite surface α-PbCSP (yellow) antibodies. DNA was stained with DAPI. Staining of the 2.34 wt parental parasite was used as a negative control. Images are de-convoluted projections of confocal stacks. BF, bright field; Scale bars in (A–C, E) 5 μm; Scale bars in (D) 10 μm.