Fig. 6: ENS cellular proliferation dynamics after benznidazole treatment of T. cruzi infections.

a Representative compressed z-stack images of transverse colon sections at 36 weeks post-infection with T. cruzi, 27 weeks post-treatment (wpt) with benznidazole (BZ), 17 weeks after EdU pulse phase. Images shown for female C3H/HeN mice that were uninfected (Control), infected with TcI-JR and (i) vehicle-administered (Infected), or treated with BZ for 20 days at 6 wpi (BZ-treated). All mice were pulsed with 6 doses of EdU between 1 and 7 weeks after withdrawal of BZ to label the progeny of cells that were proliferating during an EdU pulse (orange). Immunofluorescently-labelled Hu⁺ neuronal cell bodies (magenta) and GFAP⁺ glial cells (green). Hoechst 33342 stain shows DNA (blue). b Single z-slice merged image from a BZ-treated mouse showing intra-ganglionic EdU co-localisation events with Hu or GFAP expression (white arrows; cell 1: co-localisation with Hu at z-slice 7/18; cell 2 and 3: co-localisation with GFAP at z-slice 13/18). Orthogonal view of z-planes of the image across the x and y axes are marked by yellow lines. c, d Frequencies of EdU⁺ cells co-localising (or not) with Hu or GFAP protein expression in myenteric ganglionic (c) and peri-ganglionic (d) locations (6wpt: control n = 6, infected n = 5 and BZ-Cured n = 4; 27 wpt: control n = 4, infected n = 4 and BZ-Cured n = 7). Data also include a shorter follow-up cohort (12 weeks post-infection, 6 weeks post-treatment (wpt), 1 week after the end of the EdU pulse phase). Data are expressed as mean ± SEM, data points are for individual mice. Statistical significance was tested using Kruskal–Wallis tests (P-values < 0.1 are annotated). All confocal images were taken at 400× magnification, scale bar = 50 µm.