Figure 4 : Inhibition of apoptosis impairs blastema formation.

From: Sustained production of ROS triggers compensatory proliferation and is required for regeneration to proceed

Figure 4

(a, b) TUNEL assay and immuno-detection of active Caspase-3 were performed on 18-hpa samples (white: DMSO; grey: NS3694 5 μM). (c) The efficiency of regeneration was quantified at 72 hpa and is expressed as a percentage of control; representative fins are shown. (d) Fish were incubated in M50054 (a cell permeable inhibitor of apoptosis) at 30 μM or vehicle (DMSO) from the time of amputation to 3 dpa. The efficiency of regeneration was quantified at 3 dpa and is expressed as a percentage of the control. (e) The inhibition of the first wave of apoptosis does not affect the second wave. Fish were incubated in NS3694 (5 μM) or vehicle (DMSO) from 4 hours before amputation to 12 hpa. The fish were subsequently transferred to water until 18 hpa. The fins were collected, and the immuno-detection of active Caspase-3 was performed. Positives cells were counted in ray and inter ray 2 for each fin. No significant differences were observed. (f) Hematoxylin-stained longitudinal sections of the regenerated fin at 3 dpa with or without NS3694 treatment. In representative pictures, the dotted lines indicate amputation plane. The error bars represent the SEM (* p<0.05, **p< 0.01, ***p < 0.001).