Fig. 7

Sensitivity profiles of ISP1KD, ISP1OE, ISP2KD and ISP2OE parasites after treatment of the sand fly midgut lysate. Axenic amastigotes of WT, ISP1KD, ISP1OE, ISP2KD and ISP2OE Leishmania parasites were incubated up to 48 h in BOD incubator (25 °C) for transformation to the promastigotes. At different times of incubation (0, 4, 8, 12, 16, 20, 24, 36 and 48 h), cells were treated (for 2 h) with the gut extract (single blood-fed midgut) isolated at the corresponding period. Percent control viability of the parasites was determined from the geometric mean number of the viable parasite incubated with the midgut proteases over the number of viable parasites in identical aliquots incubated without protease treatment. The experiments were performed in triplicate and data are means ± SD from three separate experiments. An asterisk (*) denotes P ≤ 0.05 and ‘ns’ denotes non-significant as compared to WT (Student’s t-test)