Figure 3. Sirt1 inhibition causes a fin regeneration defect in larval fish
(A) Representative images of larval fins show the outcomes of the regenerative fin growth in the control and the Sirtinol + NAM treatment conditions at 3 dpa. (The white dashed line in the left panel indicates the resecting line.) The quantification in the graph on the right shows that Sirt1 inhibition by Sirtinol alone or combination of Sirtinol or FK866 with NAM significantly impairs regenerative fin growth in larval fish.
(B) The quantification shows that Sirt1 inhibition by a high concentration of NAM significantly impairs regenerative fin growth in larval fish.
(C) The schematic shows the alignment of genomic DNA sequence of the wild type and a sirt1 mutation containing a 28 bp insertion in exon 1 generated by CRISPR/Cas9, which results in a premature stop codon (indicated by an asterisk). The official allele name for this mutation is sirt1wcm18/wcm18. The agarose gel image shows that PCR of genomic DNA generates a larger band from a homozygous sirt1 mutant fish compared to wildtype (WT).
(D) Representative images (left) and the quantification result (right) show that sirt1 mutant fish fail to regenerate fins when incubated in NAM. Each dot represents fin regeneration measurement from one fish. Error bars indicate standard deviation. Student’s t test was performed to determine statistical significance. ∗p < 0.05.)
