TcUBP1-GFP ectopic expression in late-log phase epimastigotes promote full metacyclogenesis.

<p>(A) The percentage of metacyclic trypomastigotes was determined in uninduced or induced TcUBP1-GFP or GFP late-log epimastigotes cultures 5 days after addition of Tet. Values represent mean ± SD of 4 independent replicates. (B) TS-SAPA staining was assessed in metacyclic trypomastigotes derived from TcUBP1-GFP induced cultures (5 days). Images are representative of 3 independent experiments. (C) The percentage of metacyclics was determined in uninduced or induced TcUBP1-GFP or GFP late-log epimastigotes incubated in TAU-3AAG medium for 96 hs. (D) Staining for intracellular amastigotes in VERO cells 7 days after incubating with induced and uninduced TcUBP1-GFP cultures. (E) Percentage of infected VERO cells 7 days post-infection with parasites derived from induced or uninduced TcUBP1-GFP or GFP cultures. Values represent mean ± SD of 3 independent replicates. (F) Infected VERO cells with intracellular amastigotes 7 days post infection with TcUBP1-GFP cultures. Amastigotes were stained with mouse anti-<i>T</i>. <i>cruzi</i> before permeabilization (red) and with rabbit anti-<i>T</i>. <i>cruzi</i> after permeabilization (green). DNA was stained with DAPI, shown in cyan. Images are representative of 3 independent experiments. ***P< 0.001 by Anova-Tukey test. Scale bars, 5 μm. (G) Model for TcUBP1-induced morphological differentiation.