Extended Data Fig. 5: Gradual loss of early proventricular infection by cultured bloodstream form (cBSF) trypanosomes.
From: Trypanosoma brucei colonizes the tsetse gut via an immature peritrophic matrix in the proventriculus
a, Flies that had received a bloodmeal containing J10 BSFs (from murine stabilate) were dissected each day and midgut (MG) and proventriculus (PV) infections were scored before preparation for CLSM. Trypanosomes were observed in the PV from two dpi. Two experiments, n=19-22 flies per timepoint. b, Flies were given an infected bloodmeal with either BSFs (Blood), cBSFs that had been adapted to culture from the same BSF stabilate for 9 days (Adapted) and cBSFs that had been in culture for several months (Cultured). Flies were dissected at five dpi and fifteen dpi and infection prevalence was scored for both the proventriculus (PV) and the midgut (MG), or 30 dpi to check salivary gland (SG) infection. Two experiments, n=28-33 flies each timepoint. c, Retrospective analyses of salivary gland (SG) infections using either bloodstream forms (BSFs), cultured procyclic forms (PCFs) or cultured BSFs (cBSFs) for infection. Each point represents an independent experiment (with >100 flies per experiment). Life stages used for all experiments were from AnTat 1.1 90:13 strain. Mean ± SD, numbers on the plots indicate the sample size of each group, horizontal bars and p-values show statistical significance from an unpaired two-tailed t-test, normal distribution and equal variance. d, Flies were given an infected meal with cBSF T. brucei (strain AnTat 1.1 90:13 expressing mNeonGreen) and dissected every day until trypanosomes were observed in the proventriculus. In contrast to BSFs from a murine stabilate, the cultured forms do not invade the proventriculus until at least 15 dpi. The distribution is also different as very few trypanosomes can be seen in the ectoperitrophic space in the anterior midgut. Scale bars 100 µm.
