Extended Data Figure 1. Intestinal helminth infection induces ENS injury and gliosis.

(a-i) TM preparations (a-d, f-i) and cross-section (e) from the duodenum of Sox10|tdT mice immunostained for CD45 (a), CD31 (b), Pdgfra (c), c-Kit (d), Epcam (e), Sox10 (f, h) and HuC/D (f, i). Indicated are cells negative for tdT (empty arrowheads), tdT⁺ EGCs (asterisks) and neurons (arrowheads). g, h and i show single spectrum images of f. n=3. (j) Quantification of tdT⁺ cells expressing Sox10 (EGCs) and HuC/D (neurons) (n=11 field of views, representative of 3 experiments). (k) Schematic of intestinal cross-section illustrating the organisation of the EGC-network and the life-cycleof H. poly. L3 H. poly larvae penetrate the duodenum mucosa and settle in the TM, eliciting local tissue damage, inflammation and formation of granulomatous infiltrates. 10 days later they emerge as adult worms into the lumen where they mate and produce eggs. (l, m) Cross-section (l) and whole-mount view (m) of the duodenum of H. poly-infected Sox10|tdT mice (7 dpi). Schematics (top left) show the orientation of the images. Arrowheads point to H. poly settlement sites. n=4. (n, o, q, r) TM preparations from the duodenum of naïve and H. poly-infected (7 dpi n, o; 10 dpi q, r) animals immunostained for GFAP and Sox10 (n, o) and S100 (q, r).n=5. (p) Quantification (qRT-PCR) of Gfap transcripts in the TM of naïve and H. poly-infected mice (7 dpi). (n_Naïve=8, n _H.poly =6). 2 experiments. (s-u) Quantification of S100⁺ type III EGC morphology including total process length (s) (n=10), process thickness (t) (n=10) and Scholl analysis for EGC process branching (u) (n=5). 2 experiments. Two-tailed Mann-Whitney test (p, t), unpairedtwo-tailed t-test (s), Two-way ANOVA with multiple comparisons (u). Mean±SEM. Scale bars: a-i: 50 μm; l, m, n, o, q, r:100 μm, insets: 12.5 μm