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Reverse Genetics Screen in Zebrafish Identifies a Role of miR-142a-3p in Vascular Development and Integrity

Figure 8

miR-142a-3p and Cdh5 are regulated by the transcription factor Lmo2 in zebrafish.

A–F - Morpholino (MO) mediated Lmo2 knockdown induces cerebral hemorrhage phenotype in 2 dpf Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos. A,C,E - non-injected control (NIC) and B,D,F - 1 mM lmo2 MO injected zebrafish embryos. The embryos were imaged at 2.5× magnification. Arrowheads indicate the site of hemorrhage. G - cdh5 relative expression quantified by QRT–PCR in lmo2 knockdown Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos. Data collected from 4 independent experiments is represented as mean fold change ± SD. Asterisk (*) indicates p value of 0.0001 as determined by 2-tailed t-test. H - Western Blot analysis for Cdh5 in 2 dpf NIC and 1 mM lmo2 MO injected zebrafish embryos. Beta-actin was used as a loading control. I - miR-142a-3p relative expression quantified by QRT–PCR in Lmo2 knockdown zebrafish embryos. The assay was performed as described by manufacturer (QuantiMir kit, SBI, USA). The relative expression of miR-142a-3p was normalized to miR-26a. Data collected from 4 independent experiments is represented as mean fold change ± SD. Hash (#) indicates p value of 0.001 as determined by 2-tailed t-test.

Figure 8

doi: https://doi.org/10.1371/journal.pone.0052588.g008