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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

Fig. 4

circGFRA1 serves as a sponge for miR-34a. a. The levels of nuclear control transcript (U6), cytoplasmic control transcript (GAPDH mRNA) and circGFRA1 were assessed by qRT-PCR in nuclear and cytoplasmic fractions. b. The predicted binding sites of miR-34a within circGFRA1 were shown. c. Cells were transfected with miR-34a mimic or miR-34a inhibitor, qRT-PCR analysis demonstrated that the transfection was successful. d. Luciferase assay of MDA-MB-231 cells cotransfected with miR-34a mimics or miR-34a inhibitor and luciferase reporter containing circGFRA1 3′-UTR (circGFRA1 wt) or mutant construct (circGFRA1 mut). e. Cells were transfected as described, and the expression of circGFRA1 was determined by qRT-PCR. f. Cells were transfected with si-NC or si-circGFRA1, and the expression of miR-34a was determined by qRT-PCR. U6 snRNA was used as an internal control. g. MS2-based RIP assay in TNBC cells transfected with MS2bs-circGFRA1, MS2bs-circGFRA1mt, or MS2bs-Rluc (control vector) (n = 3). All the data are shown as the mean ± s.e.m., *P < 0.05 and **P < 0.01

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