V-ATPase inhibition affects regeneration and, similar to H+ efflux, has a different expression pattern depending on the amputation plane along the PD axis. (A) Inhibition of the V-ATPase using concanamycinA (concA) and fluorescein- tagged morpholinos (fluo-MO-1 and 2). (B–M) Experiments performed after proximo-distal amputation. (B–G) In situ hybridization for atp6v1e1b at 12 h (B, C), 24 h (D, E) and 48 h (F, G) after proximal (B, D, F) and distal amputation (C, E, G). (H–K) Immunohistochemical detection of Atp6v1a at 24 h (H, I) and 48 h (J, K) after proximal (I, K) and distal amputation (H, J). (L) H+ efflux pattern during regeneration after proximal and distal amputation (n = 8–10). (M) Length of Atp6v1a expression domain in proximal and distal stumps, 24 and 48 hpa. *statistical significant results (n = 3, p<0.05). hpa: hours post amputation. For each panel, n = 6, except mentioned otherwise.
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