Fig. 1

FUCCI transgenic line generation. a, b Schematic representations of FUCCI green and red constructs, respectively. FUCCI constructs were injected separately in different 1-cell stage fertilized eggs. Positive eggs were raised into adult fish, bred and screened for three generations (c). F2 FUCCI green fish were finally bred with F2 FUCCI red fish to generate double FUCCI embryos, which were used for most experiments (c). d Schematic representation of how FUCCI technology works, cells are green during S/G2/M phases, colourless between M and G1, red in G1 and G0 phases and yellow during a small portion of G2 phase. e Schematic representation of zUbiquitin-EGFP construct. EGFP expression driven by zUbiquitin promoter in Nothobranchius embryos and adult fish is shown (f). g FACS analysis of double FUCCI embryo. The scatterplot on the left shows the gating used to separate the four cell populations. The numbers indicate the percentage of cells in the four populations. The middle graph represents the intensity of the Hoechst staining as measure of DNA content of the four different populations. The graph on the right is the same as the graph in the middle but population are normalized on their own cells count rather than the total cells count. h FACS analysis of adult gonads of double FUCCI fish. The scatterplot on the left shows the gating used to separate the four cell populations. The numbers indicate the percentage of cells in the four populations. The middle graph represents the intensity of the Hoechst staining as measure of DNA content of the four different populations. The graph on the right is the same as the graph in the middle but population are normalized on their own cells count rather than the total cells count