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Fig. 4. V-ATPase function is required for the upregulation of cell proliferation
in the tail regeneration bud. Proliferating cells were labeled with
immunohistochemical staining for phosphorylated Histone H3B (H3-P). (A)
At 24 hpa, proliferating cells in the G2-M transition are randomly
distributed. (B) By 48 hpa, the proliferating cells cluster in the
regeneration bud and have largely disappeared from the caudal flank. Red
arrows indicate H3P-positive cells. (C) Incubation in concanamycin
immediately after amputation reduces proliferation in the bud; sample
proliferation data are from D (control) and D'
(concanamycin-treated) larva. A,B are oriented anterior to the left; all other
panels are oriented anterior upwards. n=8 for each condition.
(E-G') KCNK1 expression requires V-ATPase activity. The
regeneration bud normally expresses the KCNK1 channel by 24 hpa (E,E').
By contrast, larvae in which the V-ATPase was inhibited by concanamycin
(F,F') or anti-c subunit antibody (G,G') exhibit no KCNK1
expression at 24 hpa. n=10 for each condition. White arrows indicate
lack of expression; red arrows indicate positive signal (expression of
KCNK1).
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