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Fig. 7
ARF senses regenerative signals and suppresses fin regeneration. (A) Schematic of transgene expressing human ARF under the control of the human ARF promoter (left). Representative images of age- and sex-matched ARF:ARF and WT zebrafish (right; 5 months postfertilization, male). Scale bar: 1 cm. ARF:ARF fish are viable, grow to adulthood and are of normal size and patterning. (B) Immunostaining (longitudinal confocal images) for ARF in ARF:ARF transgenic fish before injury (uninjured ) and at 2 dpa. Scale bars: 50 µm. ARF is specifically expressed upon injury. The dashed line represents the amputation plane. (C) Representative images of fin regeneration at 6 dpa in WT and ARF:ARF fins (top). Scale bars: 1 mm. The dashed lines represent amputation planes. Quantification of regenerate length and area at 6 dpa in WT and ARF:ARF fins (bottom; N= 10 fins, p<0.001). The first set of bars in each graph represents the results from one transgenic line (Line 1), while the second set of bars represents the results from a second, independent transgenic line (Line 2). ARF causes marked inhibition of fin regeneration. Results are shown as mean ± standard deviation. Figure supplement 1 shows the embryonic viability of ARF transgenic lines. Figure supplement 2 shows the failure of ARF:ARF fins to completely regenerate after 15 days and even 30 days. Figure supplement 3 shows ARF immunostaining at 6 dpa, Tp53, tp53, and cdkn1a expression changes with ARF expression in WT and ARF:ARF fins at 4 dpa, fin regeneration rescue in ARF:ARF fins treated with PFTα, and EdU incorporation studies performed in WT and ARF:ARF fins. TSS: Transcriptional state site; uninj.: Uninjured; WT: Wild type. n.s.: not significant.