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Fig. 4 | Molecular Cancer

Fig. 4

From: AR–PDEF pathway promotes tumour proliferation and upregulates MYC-mediated gene transcription by promoting MAD1 degradation in ER-negative breast cancer

Fig. 4

PDEF upregulates MYC-mediated gene transcription by promoting MAD1 degradation. a Western blotting was performed to detect the expression levels of intracellular signalling components, including MEK/ERK, PI3K/AKT, MYC/MAD1, AR and EGFR, in PDEF-overexpressing SKBR-3 cells. b Western blotting was performed to detect the expression levels of MEK/ERK, PI3K/AKT, MYC/MAD1, AR and EGFR in PDEF-downregulated MDA-MB-453 cells (NS: non-specific; KD: knockdown). c Co-IP assay was performed with the anti-PDEF antibody in PDEF-overexpressing SKBR-3 cells and control vector-infected cells. The interaction between precipitated MYC and PDEF was detected using the anti-PDEF antibody. d Co-IP assay was performed with the anti-PDEF antibody in PDEF-downregulated MDA-MB-453 cells and control vector-infected cells. The interaction between precipitated MAD1 and PDEF was detected using the anti-PDEF antibody. e PDEF and MYC mRNA levels in PDEF-overexpressing SKBR-3 cells or PDEF-downregulated MDA-MB-453 cells were determined by performing RT-qPCR (left panels). PDEF and MAD1 mRNA levels in PDEF-overexpressing SKBR-3 cells or PDEF-downregulated MDA-MB-453 cells were determined by performing RT-qPCR (right panels); **P < 0.05. f Top panel: Schematic diagram of the PDEF-binding regions within the MAD1 locus. Lower left panel: Results of the direct PDEF ChIP assay followed by RT-qPCR of PDEF-downregulated MDA-MB-453 cells (black bars) or control vector-infected (white bars) cells; data are presented as mean ± SD. Lower right panel: Semi-quantitative PCR of a negative control (IgG) sample

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