Fig. 2. EDN3 signalling inhibits lineage commitment and differentiation of EPCs. Wild-type EPCs were isolated from cultures of embryonic (A-D,H-K,L-O,R,S) or postnatal (E,F,P,Q,T,U) gut and maintained in either standard (control) medium (CM; A,C,E,H,J,L,N,P,R,T) or CM supplemented with 10 nM EDN3 (B,D,F,I,K,M,O,Q,S,U). Colonies were fixed 5 (A,B,H-K,L,M) or 10 (C-F,N-Q,R-U) days after plating and immunostained for TuJ1 (A-F), MASH1 (H,I), RET (J,K), B-FABP (L,M), GFAP (N-Q) and SOX10 (R-U). In all panels, colonies were counterstained with DAPI. (G) Percentage of TuJ1⁺ cells in colonies maintained either in standard medium (CM; yellow bars) or medium supplemented with 10 nM EDN3 (CM+EDN3; green bars) 5, 7 and 10 days after plating. (V) Percentage of SOX10⁺ cells in colonies maintained in CM (yellow bars) or CM+EDN3 (green bars) 1, 5 or 10 days after plating.