A,B:
Insulin expression at NF44 is almost completely abolished (75%, n=16). C,D:
Glucagon expression at NF44 is almost entirely lost in the stomach and pancreas (67%, n=6). E,F: Expression of PDIp is almost completely lost in Staufen2 knockdown animals at NF44 (75%, n=12). G,H:
tm4sf3 gene expression is lost at NF44 (78%, n=9). I,J: Expression of hex is lost at NF44 (100%, n=11). K,L: Real-time PCR analysis of endocrine and exocrine markers in Staufen2 knockdown embryos for amylase in control and Staufen2 knockdown whole guts at NF44 and insulin expression in whole embryos at NF35. Each bar is an average of 8 individual whole tadpoles. Amylase expression is almost lost, and insulin expression is decreased by more than 2-fold. M–P: Flag-Staufen2 mRNA rescues the morpholino knockdown phenotype. M: Control NF45 whole gut stained for ptf1a mRNA showing expression in the pancreas. N: Whole gut from embryo injected with 40 ng of Staufen2 morpholino. Ptf1a expression is almost completely lost in injected embryos (n=20). O: Isolated whole gut stained for ptf1a expression from embryo injected with 40 ng of Staufen2 morpholino and 2000 pg of flag-Staufen2 mRNA. Ptf1a expression was restored in 31/45 embryos. P: In vitro transcription translation of staufen2 mRNA with either the morpholino (MO) or the mismatch morpholino (MM).