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Fig. 3 Design and evaluation of effectiveness of antisense oligonucleotides targeting pld1 translation and splicing. (A) The scheme of morpholinos designed to target pld1. (B) in vitro transcription and translation experiment testing the efficiency of MO1-pld1 to inhibit pld1 translation in a dose-dependent manner. (C) Gel electrophoresis of RT-PCR amplified pld1 fragments to monitor interference by MO2-pld1 and MO3-pld1 compared to the normal splicing of pld1. ef1a is used as a loading control. 5mmMO2-pld1: 5mismatch MO2-pld1. (D) The basal and PMA stimulated PLD activity were measured using a stable isotope method for detecting the formation of PtdBuOH by mass spectrometry. Enzyme activity is significantly reduced in MO1-pld1 injected embryos (*, p < 0.01). Data is from three independent experiments. Each condition represents the average measurement from 50 embryos at 1 dpf. Error bars depict SEM.
Reprinted from Developmental Biology, 328(2), Zeng, X.X., Zheng, X., Xiang, Y., Cho, H.P., Jessen, J.R., Zhong, T.P., Solnica-Krezel, L., and Brown, H.A., Phospholipase D1 is required for angiogenesis of intersegmental blood vessels in zebrafish, 363-376, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.