Figure 2.

Evaluation of 17-DMAG’s effectiveness against Leishmania in vitro. (A) Cytotoxicity of 17-DMAG against BMMΦ in vitro. BMMΦ were plated at a concentration of 5 × 10⁴ cells per well in a 96-well plate and incubated with different concentrations of 17-DMAG (0.024 to 50 μM) in 200 µL of DMEM low complete medium. The graph represents the median and quartiles values of the % of viable macrophages from one representative of five independent experiments carried out in triplicate, and CC₅₀ was calculated as the mean ± SE of five experiments. (B) Efficacy of 17-DMAG against axenic promastigotes of L. braziliensis. Axenic promastigotes of L. braziliensis in the exponential growth phase were plated at a concentration of 4 × 10⁵ parasites per well in a 96-well plate and incubated with various concentrations ranging from 0.001 μM to 2 μM of 17-DMAG in 200 µL of complete Schneider’s medium. The graph shows the median of the % of viable parasites treated with various 17-DMAG concentrations from one out of four independent experiments carried out in triplicate, and the IC₅₀ value is given by the mean ± SE of four experiments. (C) Effectiveness of 17-DMAG in controlling BMMΦ infection by L. braziliensis. BMMΦ were infected with L. braziliensis at a 10:1 ratio. After 24 h, the infected cells were treated with different concentrations of 17-DMAG (0.9 to 900 nM) or left untreated. Viable parasites were assessed after 6 days of differentiation. The graph represents the median and interquartile values of the number of intracellular viable parasites liberated from treated macrophages from one out of four independent experiments conducted in quintuplicate. The IC₅₀ value was calculated as the mean ± SE of four experiments. For details, see Section 2.4, Section 2.5 and Section 2.6.