Fig. 4

CEBPA negatively regulates RGCC expression. A The recognition motif of CEBPA in RGCC promoter (upper) analyzed using the JASPAR database, and the schematic illustration (down) showing the potential CEBPA responsive elements (E1). B A negative correlation between RGCC and CEBPA expression by Pearson correlation analysis using TCGA-TNBC dataset. C Luciferase reporter assays showing regulation of CEBPA regulation of RGCC promoter activity in 293 T and MDA-MB-231/LM3 cells. D ChIP assays to verify CEBPA binding affinity to the RGCC promoter in MDA-MB-231/LM3 transfected with CEBPA or control vector. E, F The regulation of CEBPA to RGCC was determined by qRT-PCR (E) and western blotting (F) in lung metastatic 4T1/LM3 and MDA-MB-231/LM3 cells. G High level of methylated CEBPA was in breast tumors than in normal tissues. H The Kaplan–Meier survival curves of BC patients with high or low methylated CEBPA. I Detection of methylation level in CpG islands of the CEBPA promoter by methylation specific polymerase chain reaction (MSP) (M: Methylation primer, U: Unmethylation primer.). (The data are presented as the mean ± SD; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001)