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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: Estrogen receptor β inhibits breast cancer cells migration and invasion through CLDN6-mediated autophagy

Fig. 3

ERβ regulates CLDN6 expression at the transcriptional level. a ERβ protein expression in the nuclear and cytoplasmic fractions of DPN-treated MDA-MB-231 cells. PCNA served as a nuclear protein control, and β-tubulin was used as a cytoplasmic protein control. b Subcellular localization of ERβ. DPN promotes the transportation of ERβ from the cytoplasm to the nucleus (white arrowheads) in MDA-MB-231 cells. Nuclei were stained with DAPI (blue) (scale bar, 50 μm). c Schematic diagram of the sequence spanning from -2000 bp to + 250 bp relative to the translational start site (TSS) of the CLDN6 gene promoter containing the (half-ERE)-(N)x-(GC-box) motif. d ChIP assay. The ERβ binding sites (− 183/− 168 bp or + 155/+ 170 bp) of the CLDN6 promoter were detected by PCR in DPN-treated MDA-MB-231 cells. e ChIP assay. The Sp1 binding sites (− 60/− 47 bp or + 192/+ 203 bp) of the CLDN6 promoter were detected by PCR in DPN-treated MDA-MB-231 cells. f The luciferase activities of DPN-treated MDA-MB-231 cells transfected with pGL3-CLDN6 and renilla luciferase reporter (pRL-TK) plasmids were detected by dual-luciferase reporter assays. Data are presented as mean ± SD. The data shown are representative results of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001

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