Characterisation of rrm2b−/− mutants—. (A) Mean velocity (mm/s) after touch stimulus of 2 dpf rrm2b+/+ (n = 8), rrm2b+/− (n = 23) and rrm2b−/− larvae (n = 12). Results are shown as mean ± SEM (statistical significance was determined with a Kruskal-Wallis test, * = adj. P < 0.05, ** = adj. P < 0.01). (B) Total displacement (mm) after touch stimulus of 2 dpf rrm2b+/+ (n = 8), rrm2b+/− (n = 23) and rrm2b−/− larvae (n = 12). Results are shown as mean ± SEM (statistical significance was determined with a Kruskal-Wallis test, * = adj. P < 0.05). (C) Peak acceleration (mm/s2) after touch stimulus of 2 dpf rrm2b+/+ (n = 8), rrm2b+/− (n = 22) and rrm2b−/− larvae (n = 12). Results are shown as mean ± SEM. (D) Representative image illustrating the morphological parameters used to assess zebrafish larval development. (E) Standard length (SL) measurements of 5 dpf rrm2b+/+ (n = 16), rrm2b+/Δ26bp (n = 48) and rrm2b−/−(n = 14) larvae. Results are shown as mean ± SEM. (F) Snout-vent length (SVL) of 5 dpf rrm2b+/+ (n = 16), rrm2b+/− (n = 48) and rrm2b−/− (n = 14) larvae. Results are shown as mean ± SEM. (G) Head-trunk angle (HTA) measurements of 5 dpf rrm2b+/+ (n = 16), rrm2b+/− (n = 48) and rrm2b−/− (n = 14) larvae. Results are shown as mean ± SEM. (H) Eye area (EA) measurements of 5 dpf rrm2b+/+ (n = 16), rrm2b+/− (n = 48) and rrm2b−/− (n = 14) larvae. Results are shown as mean ± SEM. (I) Swim bladder area (SBA) measurements of 5 dpf rrm2b+/+ (n = 16), rrm2b+/− (n = 48) and rrm2b−/− (n = 14) larvae. Results are shown as mean ± SEM (statistical significance was determined with a Kruskal-Wallis test, **** = adj. P < 0.0001). (J) Representative images of 5 dpf rrm2b+/+ larvae morphology (top panel) and 5 dpf rrm2b−/− larvae morphology (bottom panel). White arrow indicates uninflated swim bladder. (K) Relative mtDNA copy number of whole 3 dpf rrm2b+/+ & rrm2b+/− (n = 13) and rrm2b−/− (n = 14) larvae. Results are shown as mean ± SEM. (L) Relative mtDNA copy number in whole 5 dpf rrm2b+/+ & rrm2b+/− larvae (n = 50) and rrm2b−/− larvae (n = 29). Results are shown as mean ± SEM. (M) Relative mtDNA copy number in the head and abdomen of 5 dpf rrm2b+/+ & rrm2b+/− larvae (n = 21) and rrm2b−/− larvae (n = 16). Results are shown as mean ± SEM. (N) Relative mtDNA copy number in tail muscle of 5 dpf rrm2b+/+ & rrm2b+/− larvae (n = 15) and rrm2b−/− larvae (n = 12) results are shown as mean ± SEM (statistical significance was determined with a Student’s t-test, * = P < 0.05). (O) Survival of rrm2b larvae beyond 5 dpf by genotype. 5 dpf: rrm2b+/+: 25% (n = 23), rrm2b+/−: 52% (n = 48), rrm2b−/−: 23% (n = 21). 7 dpf: rrm2b+/+: 18% (n = 6), rrm2b+/−: 58% (n = 19), rrm2b−/−: 24% (n = 8). 10 dpf: rrm2b+/+: 37% (n = 15), rrm2b+/−: 53% (n = 21), rrm2b−/−: 10% (n = 4). 12 dpf: rrm2b+/+: 42% (n = 15), rrm2b+/−: 50% (n = 18), rrm2b−/−: 8% (n = 3). 14 dpf: rrm2b+/+: 16% (n = 5), rrm2b+/−: 87% (n = 27), rrm2b−/−: 0% (n = 0). 17 dpf: rrm2b+/+: 36% (n = 9), rrm2b+/−: 60% (n = 15), rrm2b−/−: 4% (n = 1). 19 dpf: rrm2b+/+: 37% (n = 12), rrm2b+/−: 63% (n = 20), rrm2b−/−: 0% (n = 0). 21 dpf: rrm2b+/+: 60% (n = 21), rrm2b+/−: 40% (n = 14), rrm2b−/−: 0% (n = 0).
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