Figure 4
Figure 4. L-leucine improved hemoglobinization and increases total numbers of erythrocytes in rps14 morphants. (A-L) Representative images of zebrafish embryos at 4 dpf stained for hemoglobin with O-dianisidine. Lateral views with head to the left, dorsal upwards (A-D), ventral (E-H), and tail (I-L). Control morphants treated with control (A,E,I) showed normal hemoglobinization, as did the control morphants treated with leucine (B,F,J). rps14 morphants treated with control (C-G-K) were severely anemic and exhibited morphologic abnormalities including short stature and lack of bronchial arch development similar to those observed in rps19 morphants. Developmental defects and anemia were alleviated after leucine treatment (D,H,L). (M) Quantification of the percentage of embryos exhibiting the phenotypes shown in panels A through L. The percentage of severely and moderately anemic rps14 morphants with leucine treatment were reduced, whereas those with normal hemoglobin/mild anemia increased (***P < .001, 2-way ANOVA with Bonferroni posttest analysis; error bars show SEM). (N) Scatter plot showing the absolute number of RFP-expressing cells per embryo. Each data point is a single embryo (**P < .01).

L-leucine improved hemoglobinization and increases total numbers of erythrocytes in rps14 morphants. (A-L) Representative images of zebrafish embryos at 4 dpf stained for hemoglobin with O-dianisidine. Lateral views with head to the left, dorsal upwards (A-D), ventral (E-H), and tail (I-L). Control morphants treated with control (A,E,I) showed normal hemoglobinization, as did the control morphants treated with leucine (B,F,J). rps14 morphants treated with control (C-G-K) were severely anemic and exhibited morphologic abnormalities including short stature and lack of bronchial arch development similar to those observed in rps19 morphants. Developmental defects and anemia were alleviated after leucine treatment (D,H,L). (M) Quantification of the percentage of embryos exhibiting the phenotypes shown in panels A through L. The percentage of severely and moderately anemic rps14 morphants with leucine treatment were reduced, whereas those with normal hemoglobin/mild anemia increased (***P < .001, 2-way ANOVA with Bonferroni posttest analysis; error bars show SEM). (N) Scatter plot showing the absolute number of RFP-expressing cells per embryo. Each data point is a single embryo (**P < .01).

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