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Fig. 4 | Genome Biology

Fig. 4

From: A novel FLI1 exonic circular RNA promotes metastasis in breast cancer by coordinately regulating TET1 and DNMT1

Fig. 4

FECR1 upregulates FLI1. a Diagram of the RNA reverse transcription-associated trap (RAT) assay. FECR1 was in situ reverse transcribed using circular RNA-specific primers in the presence of biotin-dCTP. The FECR1-interacting chromatin DNAs were isolated for library sequencing. b Location of PCR primers to detect the interaction of FECR1 at the FLI1 locus. 5′-CT, 5′-upstream control site. c Binding of FECR1 in the FLI1 locus. The FECR1 RAT-captured chromatin DNAs were amplified by PCR using primers covering the FLI1 locus. Note the binding of FECR1 in the FLI1 promoter (P1, P2). d Activation of FLI1 by FECR1. Expression of FLI1 was quantitated by qPCR using two pairs of primers that cover different regions of FLI1. Region 1, the PCR product covers exon 4 to exon 6; region 2, the PCR product covers exon 3 to exon 4. **p < 0.01 as compared with PBS and vector control groups. Both qPCR data show that the overexpressed FECR1 upregulates the linear FLI1 mRNA. e Western blot of FLI1 protein. Cells that were stably transfected with FECR1-overexpression vector, vector control, and PBS were collected for Western blotting. f Quantitation of FLI1 oncoprotein Western blot. **p < 0.01 as compared with PBS and vector control groups

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