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Fig. 5 | Molecular Cancer

Fig. 5

From: The circACTN4 interacts with FUBP1 to promote tumorigenesis and progression of breast cancer by regulating the expression of proto-oncogene MYC

Fig. 5

circACTN4 can bind to FUBP1 to upregulate MYC expression. a and b RNA pull-down experiment was performed using the specific biotin-labeled circACTN4 probe in MCF-7 cell lysates, followed by silver staining, and the protein bands were analyzed by mass spectrometry and western blot. FUBP1 was identified as a candidate protein interacting with circACTN4. c RIP assay was executed in MCF-7 cell lysates using anti-FUBP1or anti-IgG, then the enrichment of circACTN4 was detected by RT-PCR and qRT-PCR. d FISH-IF assay displayed that circACTN4 was co-localized with FUBP1 in the nucleus of BC cells (magnification, × 1000, Scale bar, 25um). e The relative expression of FUBP1 was determined in the BC cells transfected with overexpressed circACTN4 plasmid or si-circACTN4 by qRT-PCR. f Chip assay showed that FUBP1 was enriched in the FUSE upstream of MYC promoter and was not enriched in the GAPDH promoter. g and h The effects of overexpression and knockdown of FUBP1 in BC cells on the expression of MYC were detected by qRT-PCR and western blot. i The relative expression of FUBP1 in 20 pairs of BC tissues and para-cancer tissues was determined by qRT-PCR. j Pearson correlation analysis showed that the expression of FUBP1 was positively correlated with the level of circACTN4. k The relative expression of MYC in 20 pairs of BC tissues and para-cancer tissues was detected by qRT-PCR. l Pearson correlation analysis showed that the expression of FUBP1 was positively correlated with that of MYC. m The relative expression of MYC was evaluated after transfection with overexpressed circACTN4 plasmid and si-circACTN4 by qRT-PCR. n The expression of circACTN4 was positively associated with that of MYC by pearson correlation analysis. o The expressions of MYC and its downstream proteins CDK4 and CCND2 were assessed in the BC cells transfected with overexpressed circACTN4 plasmid and si-circACTN4 by western blot. GAPDH was used as the normalizing gene in the above experiments. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001

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