Exocrine pancreas proteases contribute to β-cell production. (A) Brightfield image of representative 5 dpf larva shown, boxed area indicates location of pancreas. Scale bars: 500 μm (left) and 100 μm (right). (B) Visualization of individual mCherry-expressing β-cells in control and protease mRNA injected Tg(ins:mCherry) larvae at 5 dpf. Scale bar: 10 μm. (C) β-cell count in control (n=24) and protease mRNA (n=24 for five each) injected animals at 5 dpf. (D) β-cell count in control (n=26), alms1 MO (n=29), and alms1 MO plus protease mRNA (n=26, 29, 22 and 23, respectively) injected larvae at 5 dpf. a=* compared to control, b=** compared to alms1 MO, and c=* compared to alms1 MO. (E) β-cell count in control (n=27), bbs1 MO (n=26), protease MO (n=29 for both), and bbs1 MO plus protease MO (n=26 and 29, respectively) injected larvae at 5 dpf. (F) β-cell count in control (n=29), ctrb1 mRNA (n=38), catalytically dead (D)-ctrb1 mRNA (n=40), inactivatable (I)-ctrb1 mRNA (n=31), and both catalytically dead and inactivatable (D/I)-ctrb1 mRNA (n=35) injected animals at 5 dpf. (G) β-cell count in control (n=29), alms1 MO (n=30), and alms1 MO plus: ctrb1 mRNA (n=40), catalytically dead (D)-ctrb1 mRNA (n=35), inactivatable (I)-ctrb1 mRNA (n=33), or both catalytically dead and inactivatable (D/I)-ctrb1 mRNA (n=30) injected larvae at 5 dpf. All statistics, Ordinary one-way ANOVA, error bars represent standard error of the mean, symbols represent the following significance: NS=p>0.05, *=P<0.05, **=P<0.01, ***=P<0.001, ****=P<0.0001.
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