Fig 1: BRE positively modulates the AKT pathway. Expression of molecules of the AKT signaling pathway upon BRE knockdown (A) or overexpression (B) was analyzed by western blotting. (C) The viability of BRE overexpressed Eca109 and TE-1 cells with or without 10 μM MK2206, an AKT inhibitor, treatment were analyzed using the CCK-8 assay, n = 3. Data are expressed as mean ± SEM. The two-way ANOVA was used to evaluate significant differences, n.s, not significant, **p < 0.01. (D) Eca109 and TE-1 cells with or without BRE overexpression were treated with 10 μM cisplatin for 24 h; one BRE overexpression group was simultaneously treated with 10 μM MK2206, and the percentage of apoptotic cells was analyzed by flow cytometry. (E) Quantitative data of flow cytometry, n = 3. (F) Expression of molecules of the AKT signaling upstream regulators upon BRE knockdown or overexpression was analyzed by western blotting. (G) qPCR analysis of PTEN gene expression in BRE knockdown or overexpression ESCC cells, n = 3. Data are expressed as mean ± SEM; Student's t-test was used to evaluate significant differences, *p < 0.05, **p < 0.01.
Fig 2: BRE promotes cell cycle progression of ESCC cells. Cell cycle distribution of ESCC cell lines after BRE knockdown (A) or overexpression (C) were analyzed via flow cytometry. (B,D) Quantitative data of the flow cytometry, n = 3; data are expressed as mean ± SEM. The two-way ANOVA was used to evaluate significant differences, *p < 0.05, **p < 0.01.
Fig 3: BRE promotes ESCC growth in vivo. Xenograft tumor growth curve of xenograft tumors generated via BRE knockdown in KYESE140 cells (A) or in BRE-overexpressing Eca109 cells (B), n = 6, and data are expressed as mean ± SEM; two-way ANOVA was used to evaluate significant differences, *p < 0.05, **p < 0.01. Representative photographs of immunohistochemical analysis of Ki67, p-AKT, p-mTOR, PTEN, Cleaved-Caspase3 in xenograft tumor sections formed by BRE knockdown in KYESE140 cells (C) or BRE-overexpressing Eca109 cells (E). (D,F) The immunoreactive areas in the IHC images were quantified using ImagePro Plus 6.0 software (Media Cybernetics, Silver Spring, MD). The integrated optical density (IOD) values were represented as the mean ± SEM. Student's t-test was used to evaluate significant differences, **p < 0.01.
Fig 4: BRE was overexpressed in ESCC tissues. (A) Representative images of IHC analysis of Ki67 staining in ESCC and tumor-adjacent normal tissue. (B) Representative photographs of IHC analysis with high, medium, and low intensities of BRE staining in ESCC and tumor-adjacent normal tissue. (C) Immunohistochemistry score of BRE in 50 pairs of ESCC and peri-tumor tissues. (D) Percentage of samples with high, medium, and low BRE protein levels in 50 pairs of ESCC and peri-tumor tissues. (E) BRE protein levels in eight ESCC (T) and peri-tumor tissues (P); (F) BRE gene expression levels in eight paired ESCC and tumor-adjacent tissues. (G) BRE protein levels in five ESCC cell lines and normal esophageal epithelial cell lines HEEC. Data are expressed as mean ± SEM values; paired Student's t-test was used to evaluate significant differences, **p < 0.01.
Fig 5: BRE promotes ESCC cell proliferation. (A,B) The viability of ESCC cell lines after BRE knockdown or overexpression was determined using the CCK-8 assay. Data are expressed as mean ± SEM; two-way ANOVA was used to evaluate significant differences, **p < 0.01; (C,D) Representative photographs of EdU-incorporated cells after BRE knockdown or overexpression. (E) Representative photographs of clone formation assays in different ESCC cell lines after BRE knockdown or overexpression.
Supplier Page from Abcam for Anti-BRCC45/BRE antibody [EPR11858]