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Fig. 4
ARF suppresses fin regeneration.(A) Schematic of heat shock regimen. An initial hour long, 37°C heat shock is delivered 3 hr prior to amputation (0 dpa) and then every 6 hrs thereafter for 6 days. Regenerates are then assessed (top) or fins are reamputated (0 dpa) and allowed to regenerate in the absence of heat shock for 6 days (bottom). (B) Quantification of regenerate length and area at 6 dpa in WT and hs:ARF fins exposed to the heat shock regimen (left; N = 40 fins representing multiple different transgene insertions, p<0.001). Representative images of fin regeneration at 6 dpa in WT and hs:ARF fins exposed to the heat shock regimen (right). (C) Quantification of regenerate length and area at 6 dpa in reamputated hs:ARF fins not exposed to heat shock (left; N = 40 fins, p>0.05). Representative image of fin regeneration at 6 dpa in a reamputated hs:ARF fin not exposed to heat shock (right). The dashed lines represent amputation planes. Scale bars: 1 mm. Results are shown as mean ± standard deviation. Figure supplement 1 shows ARF and Tp53 immunostaining at 4 dpa, and tp53 and cdkn1a expression changes with ARF expression. It also shows regeneration at 14 dpa after heat shock was discontinued at 6 dpa. hs: Heat shock; WT: Wild type.n.s.: not significant.