Leishmania HASP and SHERP Genes Are Required for In Vivo Differentiation, Parasite Transmission and Virulence Attenuation in the Host
Fig 5
Osmotaxis and PSG detection assays.
A) The osmotaxis assays analysed the capacity of each parasite line tested to follow attractant gradients in vitro. Attractant coefficients were calculated (dividing parasite numbers attracted by sucrose by parasite numbers in attractant free capillaries). No significant differences in comparison to the parental line (FVI) were shown by parametric one-way ANOVA (P = 0.206). B) Light microscopic images of compressed infected thoracic midguts (TMG) dissected from FVI and cDNA16 dKO infections. Parasites immobilized in PSG spread around the ruptured stomodeal valve (SV), allowing visualisation of the PSG, as shown for FVI. In TMGs lacking PSG, this is not observed on squeezing the midgut, as shown for cDNA16dKO. The images were marginally adjusted for contrast and sharpness to improve visibility. C) PSG extracts from 10 infected midguts per parasite line were dot-blotted on activated nitrocellulose membranes and probed with the LT15 antibody. PSG was only detected in FVI and cDNA16 sKI. LT15 was blotted onto the nitrocellulose membrane as a control for detection by the appropriate secondary HRP antibody.