Ballari_et_al_Supporting_Information_01_T_cruzi_growing_curves.pdf

T cruzi growing curves: effect of the proline analogues on the proliferation of the epimastigote form of T. cruzi

Growth curves, dose response curves, and IC50s obtained for treatments of different strains of T. cruzi epimastigotes with variable concentrations of proline analogues, as well as growth curves for washout experiments on T. cruzi epimastigotes.

Exponentially proliferating T. cruzi epimastigotes (2.5x106 cells/mL) were seeded in 96-well plates and further treated with different concentrations of each analogue (concentration range from 25 to 250 µM) or untreated (negative control). As a positive control, a combination of rotenone (60 µM) and antimycin (0.5 µM) was used. The plates were incubated at 28 °C for 8 to 10 days. Cell proliferation was spectrophotometrically measured by recording the absorbance at 620 nm every 24 h for 8 to 10 days. The absorbance was transformed into cell density values (cells/mL) using a linear regression equation that was previously obtained under the same conditions. Data from the mid-exponential growth phase were used to determine the concentration of the analogues that inhibited 50% of the parasite proliferation (IC50). For this, we explored classic dose-response sigmoid functions to find the best fit to the obtained data. Each experiment was made in triplicates, and the results presented correspond to the average of three independent experiments.