FIG 4.

Ergosterol biosynthesis pathway. Sterols that increased, decreased, or were detected but with no change as a response to miltefosine exposure are shown on the pathway (L. donovani axenic amastigotes with 5-h drug exposure in blue, L. donovani axenic amastigotes with 24-h drug exposure in red, L. major wild-type promastigotes in green). In all cases, trends were seen for both low and high concentrations of miltefosine treatment. Chromatographic peaks shown for ergosterol, cholesterol, 5,7,24(28)-ergostatrienol, and 5-dehydroepisterol in L. major wild type (black trace) and ΔLCB2 mutants (red trace) to highlight the differences in sterol profiles between them. 5,7,24(28)-Ergostatrienol and 5-dehydroepisterol share the same logP, and therefore, it is not possible to distinguish to which peak (9.8 to 9.9 min or 10.2 to 10.3 min) these sterols correspond.