Skip to main content
. 2018 May 16;9:172. doi: 10.3389/fgene.2018.00172

Figure 2.

Figure 2

Chicken circSVIL interacted with miR-203. (A) Chicken circSVIL is drive from exon 6 to 14 of SVIL gene and harbored four miR-203 binding sites. The yellow oval indicates perfected 5′ seed pairing binding site of miR-203, and blue ovals indicate other three potential binding sites of miR-203. (B) The perfected 5′ seed pairing binding sites of miR-203 in circSVIL. Mut indicates the mutation sequences of binding sites. (C) DF-1 cells were co-transfected with wild-type (WT) or mutant (MUT) luciferase reporters with miR-203 mimic or control duplexes. The relative levels of firefly luminescence normalized to Renilla luminescence are plotted. Error bars represent S.D. (n = 6). (D) Luminescence was measured 48 h after co-transfected with the luciferase reporter and miRNA mimics or negative control (NC) and with circRNAs overexpression vector or empty vector (EV). The relative levels of Renilla luminescence normalized to firefly luminescence are plotted. Error bars represent S.D. (n = 6). (E) Immunoprecipitation of AGO2 from myoblasts co-transfected with miR-203, miR-206 or miR-NC, and circSVIL overexpression vector. The expression level of circSVIL was quantified by qRT–PCR and normalized by GAPDH, and the fold change of immunoprecipitate/input are plotted. Error bars represent S.D. (n = 3). (F) RNA pull-down from the myoblast lysates after transfection with 3′ end biotinylated miR-1a, miR-206, or miR-203 control. The expression level of circSVIL was quantified by qRT–PCR, and fold enrichment in the streptavid in captured fractions are plotted. Error bars represent S.D. (n = 3). Student's t-test (two-tailed) was performed for data analysis. *P < 0.05; **P < 0.01; a,bP < 0.05; A,BP < 0.01.