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

Figure 7

In zebrafish, miR-142a-3p regulates cdh5.

A - cdh5 relative expression quantified by QRT–PCR upon overexpression of miR-142a-3p. Total RNA was isolated from 2 dpf non-injected control (NIC) and 10 µM miR-142a-3p duplex injected Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos. 2 ug of total RNA was used for preparing cDNA. Beta-actin was used as an internal control. Data collected from 4 independent experiments is represented as mean fold change ± SD. Asterisk (*) indicates p value of 0.001 as determined by 2-tailed t-test. B - Western Blot analysis for Cdh5 protein in zebrafish embryos using previously tested antibody directed against human VE-cad (110 kDa) [13] in non-injected control (NIC) and 10 µM miR-142a-3p duplex injected 2 dpf zebrafish embryos. Beta-actin was used as a loading control. C - cdh5 relative expression quantified by QRT–PCR upon downregulation of miR-142a-3p. Total RNA was isolated from 3 dpf non-injected control (NIC) and 200 µM miR-142a-3p MO injected Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos. 2 ug of total RNA was used for preparing cDNA. Beta-actin was used as an internal control. Data collected from 4 independent experiments is represented as mean fold change ± SD. Hash (#) indicates p value of 0.03 as determined by 2-tailed t-test.

Figure 7

doi: https://doi.org/10.1371/journal.pone.0052588.g007