Journal: Nature Communications
Article Title: Endothelin-3 and T-type Ca 2+ channels drive enteric neural crest cell calcium activity, contractility and migration
doi: 10.1038/s41467-025-68121-5
Figure Lengend Snippet: All experiments in ( a – c ) are performed at E11.5 at the ileo-cecal junction. Error bars are ±SD. a CA change relative to control before drug application for EDTA (2 mM, n = 6), GdCl3 (100 µM, n = 6), 2-APB (100 µM, n = 5), ryanodine (10 µM, n = 5). b CA change for nifedipine (Nif, 10 µM, n = 5), ruthenium red (RuR, 100 µM, n = 6), Z944 at 1 µM ( n = 7), 10 µM ( n = 7), 50 µM ( n = 9), ascorbic acid (AA, 1 mM, n = 13) and SAK3 at 1 µM ( n = 6), 10 µM ( n = 6). c CA change for NPPB (100 µM, n = 11), NFA (50 µM, n = 9) and DIDS (500 µM, n = 5). Two-tailed p -values for (a-c) are calculated from the Student t-test. Sox10 and CaV3.1 ( d ), CaV3.2 ( e ) and CaV3.3 ( f ) IHCs of ENCCs migrating from an intestinal explant on a substrate. All three T-type Ca 2+ channels are expressed by ENCCs, but not by the surrounding mesenchymal cells. The small bright green dots are AF647 µ-beads fluorescing in the same range as the T-type Ca 2+ channel secondary antibody. They were added in the overlying collagen gel for another experiment (see Fig. ).
Article Snippet: After fixation, blocking and permeation, samples were incubated for 1 day in 1:200 mouse Anti-SOX10 and 1:100 rabbit anti CaV3.1 (Alomone Labs #ACC-021) or 1:100 rabbit anti CaV3.2 (Alomone Labs #ACC-025) or 1:100 rabbit anti CaV3.3 (Alomone Labs #ACC-009) for 24 h, washed 3 times, incubated in secondary 1:500 anti-mouse Cy3 and 1:500 anti-rabbit AF647 antibody for another day, washed and imaged.
Techniques: Control, Two Tailed Test