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. 2015 Apr 27;10(4):e0125063. doi: 10.1371/journal.pone.0125063

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© 2015 Valigurová et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 9 — A-B. Actin labelling with a medium intensity in a trophozoite (PFA fixation). CLSM, IFA (A) and CLSM in a combination with transmission LM, IFA/DAPI (B). B represents a single median optical section. C. Actin labelling in a macrogamont treated with 30 μM JAS for 7 hours (PFA fixation). Note the increased accumulation of parasite actin (FITC) organised in longitudinal bands exhibiting strong fluorescence and strong F-actin (TRITC) labelling with a diffuse character. CLSM, IFA/phalloidin-TRITC. D. A gamont exhibiting a more diffuse actin (FITC) labelling of medium intensity after treatment with 10 μM cytochalasin D for 9 hours (PFA fixation). The F-actin (TRITC) labelling of the parasite did not change significantly. CLSM, IFA/phalloidin-TRITC. E. Very strong myosin (TRITC) labelling restricted to the PS and host tissue (PFA fixation). CLSM, IFA/DAPI. F. Strong spectrin (FITC) labelling of the PS in a macrogamont (PFA fixation). CLSM, IFA/DAPI. Single median optical section. G. Labelling of α-tubulin (FITC) of strong intensity in a young microgamont (PFA fixation). CLSM, IFA/DAPI. H-I. A trophozoite (fixed in ice-cold methanol) exhibiting a labelling of medium intensity for α-tubulin (FITC) and very strong intensity for myosin (TRITC). CLSM, IFA/DAPI. J. Labelling of α-tubulin (FITC) and myosin (TRITC) in an early trophozoite treated for 7 hours with 10 μM oryzalin (fixed in ice-cold methanol). The fluorescence signals for both antibodies did not change significantly. CLSM, IFA. K-L. Localisation of α-tubulin (FITC) and myosin (TRITC) in an individual (probably a young microgamont) treated with 30 μM oryzalin for 3 hours (fixed in ice-cold methanol). The fluorescence signal for tubulin became very weak, while it remained very strong for myosin. CLSM, IFA/DAPI. M. Co-localisation of α-tubulin (FITC) and F-actin (TRITC) in a macrogamont treated for 7 hours with 10 μM oryzalin (PFA fixation). CLSM, IFA/phalloidin-TRITC. N-O. Labelling of α-tubulin (FITC) and F-actin (TRITC) in a maturing trophozoite treated for 3 hours with 30 μM oryzalin (PFA fixation). CLSM, IFA/phalloidin-TRITC/DAPI. In both the preparations (M-O), there was almost no fluorescence signal for α-tubulin, while the F-actin labelled with a strong intensity. arrow—tail of the PS, asterisk—parasite attachment site, black arrowhead—PS, h—host tissue, n—parasite nucleus, white arrowhead—parasite pellicle.