Rna oligonucleotides

The full length of gga-miR-451 potential promoter region and 10 deletion fragments were amplified by PCR and sub-cloned into the KpnI/HindIII sites of the pGL3-basic vector (Promega, Madison, WI, USA). Seven promoter constructs containing site-specific mutations were generated by overlap-expression PCR. To construct transcription factors AhR and Arnt expression vectors, CDS of AhR and Arnt were amplified from cDNA derived from DF-1 cells and cloned into pCDNA3.1 (p-AhR, p-Arnt). The expression vectors were double-digested with HindIII and KpnI, and then cloned into the pCMV-C-HA vector (Beyotime, Shanghai, China). The recombinant plasmids were named pCMV-C-HA-AhR and pCMV-C-HA-Arnt, respectively. The primers are described in Table 1. All plasmids were verified by sequencing.
The sequences of all of the primers used in this study are shown in Table 1. RNA oligonucleotides were designed and synthesized by RIBOBIO (Guangdong, China) and are shown in Table 2.

miR-126 mimics and miR-126 inhibitor and negative control RNA-oligonucleotides were purchased from Guangzhou Ribobio, Guangzhou, China. Electroporation of RNA-oligonucleotides into EPC-derived exosomes was performed using 4D-Nucleofector System Manual (Lonza, Basil, Switzerland) according to the manufacture’s instruction. Briefly, 10 μg exosome was precipitated and resuspended in 90 μl electroporation buffer of Lonza Cell Line Nucleofector Kit (Lonza, Basel, Switzerland), then mixed with 10 μl miR-126 mimics, miR-126 inhibitor or negative control before the mixture was transferred into cold 0.2 cm electroporation cuvettes and electroporated at 150 V/100 μF. After remove the free-floating miRNA, exosomes were re-isolated using ultracentrifugation. The final pellet (exosome) was resuspended in PBS, and stored at − 80 °C. RT-qPCR was performed to investigate the expression level of miR-126 in exosomes.