FIGURE

Fig. S4

ID

ZDB-FIG-160304-29

Publication

Ye et al., 2016 - An insulin signaling feedback loop regulates pancreas progenitor cell differentiation during islet development and regeneration

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Fig. S4

Knockdown of insulin via insulin-a morpholino (insaMO). (A) Diagram illustrating insa pre-mRNA structure in wild type and insaMO-injected cells. insaMO targets the exon 2-intron 2 boundary of preproinsulin-a mRNA and generates two nonsense alternative splicing products. Primers complementary to exon 1 and exon 3 amplify the differentially spliced cDNA products. (B) Agarose gel showing PCR amplified insa cDNA products; mRNA was extracted from control and insaMO-injected embryos at 2 days post fertilization (dpf). Note that a single PCR product with size of 420 bp was detected in control embryos but two aberrant PCR products—520 and 360 bp—were detected in insaMO-injected embryos. β actin was used as internal control. (C) The PCR products in control and insaMO-injected embryos were extracted, cloned into pJet vector, and sequenced. insaMO-injected embryos showed two aberrant splice products: Product 1 included intronic sequence; Product 2 showed partial deletion of exon 2. (D) Predicted peptide sequence of control and two aberrant splice products. (E) No overt gross embryonic phenotypes resulted from injection of increasing doses of insaMO ranging from 2 ng to 8 ng.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 409(2), Ye, L., Robertson, M.A., Mastracci, T.L., Anderson, R.M., An insulin signaling feedback loop regulates pancreas progenitor cell differentiation during islet development and regeneration, 354-69, Copyright (2016) with permission from Elsevier. Full text @ Dev. Biol.