Figure 6

Figure 6 Pharmacological inhibition of ATR in adults affects regeneration.

(A) Experimental design of regeneration assay in adult zebrafish. ATR inhibitor IV is administered to adult fish tanks immediately after tail fin amputation. Regenerative capacity (arrowhead and line) and ATR/ATM p-substrate levels are tested at 24 and 48 h post amputation (hpa), compared with controls treated with DMSO. The caudal peduncle is indicated in a blue dotted line. On the right, representative pictures of caudal fin regeneration after 48 h post-treatment. Scale bar, 1 mm. (B) Representative WB for detection of phosphorylated ATR/ATM substrates in regenerating blastema of tert mutant adult fish. (C) Quantification of the WB. The intensity of the whole lane was used. n = 3 experiments. Exact p values are p = 0.0348 for DMSO vs 10 μM and p = 0.0072 for DMSO vs 50 μM. (D) Regeneration area of the caudal fin in adults at 24 hpa. n = 3. Each dot represents a biological replicate. Exact p value for control vs ATR inhibitor is p = 0.0053 in tert^−/−. (E) Regeneration area of the caudal fin in adults at 48 hpa. n = 3. Each dot represents a biological replicate. Exact p value for control vs ATR inhibitor is p = 0.0013 in tert^−/−. All data were mean + s.e.m. *P < 0.05, **P < 0.01, and *** P < 0.001 for one-way ANOVA plus Sidak´s post hoc test. When exact p values are not indicated is because GraphPad Prism software does not show it. Source data are available online for this figure.