Fig. 2

Large Leishmania-PV in B-1P cells is independent of the LYST/Beige gene. B-1P cells and BMM were infected with stationary-phase promastigotes of L. amazonensis for 2 h, washed, and incubated for either 24–96 h. (A) Phase contrast images showing higher magnifications of Leishmania-containing PV in B-1P cells or BMM. Schematic representation of Leishmania-PV diameter measurement. Scale bar, 10 μm. (B) Quantification of the diameter of Leishmania-PV in B-1P cells and BMM, after 96 h. The values represent the mean of 80 independent PV measurements. Results correspond to the mean ± SD of three independent biological assays. ***, p < 0.001. (C) Level of LYST/Beige transcripts in B-1P cells and BMM analyzed by qPCR. Results correspond to the mean ± SD of three independent biological assays. ***, p < 0.001 (Student’s t-test). (D) Representative phase contrast images showing the effect of vacuolin-1 (1 µM) in the Leishmania-PV (white arrows) from B-1P cells and BMM, when compared to images in (A). Scale bar, 10 μm. (E) Quantification of Leishmania-PV diameter in B-1P cells and BMM, untreated (ctr) or pretreated cells with vacuolin-1 (vac-1) and measured after 96 h of infection. The values represent the mean of 50 independent PV measurements, in each condition. Results correspond to the mean ± SD of three independent biological assays. *, p < 0.05; ***, p < 0.05 (one-way ANOVA). (F) Quantification of the infection index from the assay in (D). Data correspond to the mean ± SD of three independent biological assays. *, p < 0.05 (one-way ANOVA).