|
Fig. 3 Zebrafish models of rpl17 ablation display anemia and craniofacial patterning defects. (A) Representative lateral views of gata1:dsRed larvae imaged at 3 days postfertilization (dpf). Fluorescent signal, indicative of erythroid precursors and erythrocytes, was quantified in a consistently sized region of interest (red box) located posterior of the cloaca on the ventral side of controls, F0 mutants, and MO-injected larvae. Anterior, left; posterior, right. (B) Quantification of dsRed+ cells (erythroid cells) in the region of interest (see panel A) in F0 mutant or morphant larvae at 3 dpf. n = 20–40 larvae/batch, repeated at least twice. (C) Representative ventral views of –1.4col1a1:egfp larvae imaged at 4 dpf. Fluorescent signal was assessed for cartilage patterning defects by measuring the angle of the ceratohyal (ch) cartilage (dashed lines). Ceratobranchial (cb) arches were also dysplastic and reduced in number compared with controls. Anterior, left; posterior, right. (D) Quantification of the ch angle in F0 mutant and morphant larvae at 4 dpf; n = 16–32 larvae/batch, repeated at least twice. mRNA encodes predicted proteins p.A73_K105del and p.(T151Rfs*25) corresponding to Family 1 and Family 2, respectively. mRNA coding for p.Q56L is present in public databases (rs753489644; gnomAD browser), and scores as a benign variant in this assay. In panels B and D, ends of the whiskers are set at 1.5 times the interquartile range (IQR) above the third quartile and below the first quartile, respectively. Black dots, minimum and maximum outliers; *P < 0.05; **P < 0.01; **** P < 0.0001; (Kruskal-Wallis with Dunn’s multiple comparisons test).