A) Exemplary DNA histogram of two diploid larvae pooled in one sample. Standard size G1-peak = 1.44 pg, estimated size G2-peak = 2.85 pg, ratio G2 / G1 = 1.98, RCS value = 6.96. B) Exemplary DNA histogram of one diploid and one triploid larva pooled in one sample. Standard size G1-peak 2n = 1.44 pg, estimated size G1-peak 3n = 2.15 pg, ratio G1-3n / G1-2n = 1.49, RCS value = 2.11. In both A) and B) the dashed grey line is the initial model estimate and the red line is the fitted model. Colored circles represent matching cell cycle phases derived from a diploid or triploid larva. Peaks C, D, and E represent endopolyploid cell populations.

A) Cell number of diploid and triploid larvae in pooled FCM samples. Paired t-test, *** p < 0.001, n = 50. B) Ratio of cells in G1 and G2 phase, calculated as G2/G1*100 for each diploid and triploid larva. Welch’s t-test, *** p < 0.001, n = 50.

A) Microscopic picture of diploid erythrocytes (40x). B) Microscopic picture of triploid erythrocytes (40x). C) Zoomed in version of picture A, showing automatic cell area detection of diploid cells. D) Zoomed in version of picture B, showing automatic cell area detection of triploid cells. Note that only non-touching cells are analysed. E) Estimated cell volumes of diploid and triploid erythrocytes. Two sample t-test, *** p < 0.001, n = 40.

A) rps11, ribosomal protein S11. B) actb1, actin, beta 1. C) eef1a1l1, eukaryotic translation elongation factor 1 alpha 1, like 1. D) polr2d, polymerase (RNA) II (DNA directed) polypeptide D. E) rpl13a, ribosomal protein L13a. F) tbp, TATA box binding protein. Expression values for each gene are normalized using a combined index of the relative quantity of the other five housekeeping genes. Values are presented as means with standard deviations. Asterisks in titles indicate a significant difference in expression levels without an interaction with dpf. ANOVA, * p < 0.05, n = 3x3 larvae per ploidy level per day.

Expression values are normalized using a combined index of the relative quantity of the housekeeping genes eef1a1l1 and actb1. Values are presented as means with standard deviations. ANOVA, p = 0.39, n = 3x3 larvae per ploidy level per day.

A) Body length of diploid and triploid larvae. Two sample t-test, p = 0.96, n = 128. B) Representative picture of diploid larva at 5 dpf. C) Representative picture of triploid larva at 5 dpf.

A) Development of diploid larvae, n = 82. B) Development of triploid larvae, n = 55. In both A) and B) the dashed grey line is the reference x = y. Solid grey lines are the hours post fertilization at which the embryos and larvae were staged, namely: 6, 24, 30, 48 and 72 hours. ANOVA, p = 0.47.

A) Swimming velocity of diploid and triploid larvae at the first startle stimulus. B) Swimming velocity of diploid and triploid larvae at the second startle stimulus. C) Swimming velocity of diploid and triploid larvae at the third to tenth startle stimulus. ANOVA, *** p < 0.001, n = 124.