Fig. 4: Accelerated differentiation activated by the expression of three peptidases is mediated via the QS signalling pathway.
a Infection profile for mice infected with parasite lines ectopically expressing each peptidase (Tb927.11.2500, Tb927.11.12850 or Tb927.8.7020) under doxycycline regulation with the QS signalling pathway disrupted (ΔRBP7A/RBP7B). In each case, six mice were infected (biologically independent samples), with three being provided with doxycycline to induce ectopic peptidase expression (uninduced, blue; induced, red). In all cases, the parasitaemia (mean ± SEM) progressed unchecked due to the lack of transduction of the QS signal caused by deletion of RBP7A/B. b Confirmation of the inducible expression and extracellular release of each peptidase in an ΔRBPA/B background. Parasites were harvested from the infections shown in a, with three mice being induced for ectopic peptidase expression and three being uninduced as biologically independent samples. Detection of the stumpy marker PAD1 demonstrated equivalent stumpy generation in the parasites regardless of the inducible expression of the peptidase. Stringent doxycycline-regulated expression is observed in each case. The loading control for the cell pellet was EF1-alpha. Parasites were not successfully purified from one of the mice in the +Dox group at the end of the experiment due to a failure with the DEAE resin. c Quantitation of PAD1 expression on day 6 post infection when parastaemias exceeded 1 × 109/ml. PAD1 immunofluorescence assays were based on three biologically independent infection profiles represented in a and analysed by western blotting represented in b. Source data are provided as a source data file.
