Cell cycle inhibition and EOM regeneration. (A) Bortezomid is a proteasome inhibitor that blocks cell cycle [66, 67]. Fish were treated with 5 μM bortozemid by 2000X dilution of a 10 mM DMSO stock in fish water, same DMSO concentration was used in control group. Cell proliferation at 24 hpi was analyzed by intraperitoneal EdU injection as described before. (B) 5-fluorouracil (5-Flu) is a pyrimidine analog that blocks cell cycle through irreversible inhibition of thymidylate synthase . Fish were injected with 10 mM 5-Flu diluted in PBS, PBS injections were used as control. Cell proliferation at 24 hpi was analyzed by intraperitoneal EdU injection as described. Both treatments (bortezomib, A, or 5-Flu, B) effectively blocked cell cycle progression (no EdU staining) in the regenerating muscle. Note that the mesenchymal transition did not progress and the injured muscle retained its typical sarcomere architecture, as evidenced by DIC microscopy. Pictures are representative examples of 5 fish per group.
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