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Fig. 5 | Parasites & Vectors

Fig. 5

From: Host cell invasion and oral infection by Trypanosoma cruzi strains of genetic groups TcI and TcIV from chagasic patients

Fig. 5

Involvement of gp82 and protein kinase C in host cell invasion by T. cruzi metacyclic forms. a HeLa cells were incubated with parasites for 1 h in full-nutrient D10 or in PBS++. After fixation and Giemsa staining, the number of intracellular parasites was counted in a total of 250 cells. Values are the means ± SD of three independent assays performed in duplicate. The difference in the invasion rate in D10 and PBS++ was significant for strains 515, 588 and 1522 (***P < 0.005), 28 (**P < 0.01), 1434 (*P < 0.05). b Metacyclic forms were incubated with HeLa cells in PBS++, in absence or in the presence of rGP82, the recombinant protein based on gp82, or GST as control. After 1 h, the cells were processed as in (a) for intracellular parasite counting. Values are the means ± SD of three independent assays performed in duplicate. The recombinant gp82 significantly inhibited invasion by strains 28 (*P < 0.05), 515, 588, 1522 and 1434 (**P < 0.005). c HeLa cells were pretreated for 30 min with PMA and then incubated with metacyclic forms in PBS++. After 1 h incubation, the cells were processed as in (a) for intracellular parasite counting. Values are the means ± SD of three independent assays performed in duplicate. Pretreatment with PMA significantly inhibited invasion of strains 28, 515, 1522 (**P < 0.0001), 588 and 1434 (*P < 0.001). d Hela cells, untreated or pretreated for 30 min with PMA in D10, were incubated in PBS++ for 30 min and processed for confocal immunofluorescence for visualization of lysosomes (red) and nucleus (blue) under 60X 0bjetive. Note the perinuclear accumulation of lysosomes in PMA-treated cells. Scale bar = 20 μm

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