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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: The role of c-Myc-RBM38 loop in the growth suppression in breast cancer

Fig. 5

RBM38 directly binds to multiple regions in the 3′-UTR of c-Myc mRNA. a Schematic presentation of c-Myc transcript and the location of probes used for REMSA. AREs were shown in shaded boxes. The mutant probes were also stated briefly. b Probes B and C, but not probe A, were associated with RBM38. REMSA was performed by mixing probes A, B, C with RBM38 protein respectively. p21 cold probe, derived from 3′-UTR of p21 mRNA, was used as competitive probe. Competition assay was performed by the addition of an excess amount of p21 cold probe to the reaction mixture containing RBM38 protein and biotin-labeled probe. The arrow indicated RNA-protein complexes (RPCs). c Schematic representation of the luciferase plasmid with 3′-UTR-A, 3′-UTR-B 3′-UTR-C and 3′-UTR-D whose sequences were identical to probes A, B, C and p21 cold probe respectively. d, e The luciferase activity for the reporter carrying 3′-UTR-B or -C was repressed by RBM38. MCF-7 and ZR-75-1 cells with RBM38 overexpression lentivirus (RBM38) and the control (NC) were transfected with pGL3 control reporter or pGL3 reporter carrying 3′-UTR-A, 3′-UTR-B and 3′-UTR-C and 3′-UTR-D, separately. The fold changed in relative luciferase activity was a product of the luciferase activity induced by RBM38 divided by that induced by the NC. these data were calculated from three separate experiments and performed as mean ± SEM, *P < 0.05

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