FIG. 5.

Overexpression of LbRos3 increases LbMT trafficking to the PM and MLF uptake and sensitivity in L. braziliensis promastigotes. (A) Overexpression of LbRos3 and LdRos3-GFP in crude membrane fractions of L. braziliensis LH-2419 promastigotes was detected by immunoblotting with anti-LdRos3 antibodies. The positions of molecular mass markers (kilodaltons) are indicated on the left. (B) L. braziliensis LH-2419 promastigotes transfected with the LbRos3 or LdRos3-GFP construct expressed increased levels of LbMT and LbRos3 or LdRos3-GFP, at the PM. Western blot assays representative of at least three independent experiments are shown. The positions of molecular mass markers (kilodaltons) are indicated on the left. (C) [¹⁴C]MLF internalization after 60 min in L. braziliensis LH-2419 promastigotes overexpressing LbRos3 and LdRos3-GFP. Bars shown are the mean ± standard deviation of three independent experiments, and results are expressed as a percentage of the internalization by wild-type L. donovani. (D) MLF sensitivity after 72 h of culture in L. braziliensis LH-2419 parasites overexpressing LbRos3 and LdRos3-GFP. The EC₅₀s shown are the mean ± standard deviation of three independent experiments.