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Fig. 3 | Molecular Cancer

Fig. 3

From: CircMET promotes tumor proliferation by enhancing CDKN2A mRNA decay and upregulating SMAD3

Fig. 3

The transcription of circMET is enhanced by NONO-TFE3. (A) ChIP assay and real-time PCR were used to determine the binding affinity of NONO-TFE3 to MET promoter regions in UOK109 and 786-O cells. Real-time PCR with IgG was performed as the control. (B) Schematic summary of the dCas9-gRNA-guided ChIP (upper); Western blot was performed after dCas9-gRNA-guided ChIP (lower). (C-D) HEK293T cells were co-transfected with MET promoter-luciferase truncations and NONO-TFE3 plasmids, and the luciferase activity was determined using a dual luciferase reporter assay after 48 h. (E) Dual luciferase assay of HEK293T cells co-transfected with firefly luciferase constructs containing the wild-type or mutant NONO-TFE3 potential binding sites of MET promoter and NONO-TFE3 plasmids were performed. (F) The protein level of NONO-TFE3 and the circMET expression levels were detected after transfection with sh-TFE3 for 48 h. (G-H) Cell viability of UOK109 and 786-O cells was determined using CCK-8 assays after transfection for 48 h. (I-K) A colony formation and tumor sphere formation assay were used to determine the colony and tumor sphere formation ability of UOK109 and 786-O cells co-transfected with indicated lentivirus. (L) Cell cycle analysis was performed using flow cytometry in cells transfected with indicated lentivirus. The data are presented as the mean ± SD, n.s. = non-significant, **P < 0.01, ***P < 0.001

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