Figure 4.

Effects of HSP90 phosphorylation site mutations on the in vitro infectivity (A–F). Recombinant parasites were first pre-treated with RAD for 48 h at IC₉₀ and then used to infect bone marrow-derived macrophages (MOI 10:1). After 4 h, free parasites were removed and the infected macrophage cultures were further incubated at 37 °C and 5% CO₂ for 44 h. Genomic DNAs were isolated from free and attached macrophages and subjected to a duplex qPCR against mouse and Leishmania actin DNA (Bifeld et al., 2016). The median parasite:host DNA ratio was then compiled. Abbreviations: (n.s.): not significant, *p ≤ 0.05, **p ≤ 0.01 in a one-tailed, paired Student’s t test (A), or in a two-tailed U-test (B–F); n = 6.