Fig 1: IHC scoring of ERa, ERß, and GPER staining of the 12 tissue samples from our cohort. Before-and-after graphs show (A) ERa (anti-ERa antibodies, 1D5, Dako, Cat. #: M7047, lot 1: 00034057, lot 2: 20015818), (B) ERß (anti-ERß antibodies, ab3576, Abcam, Cat. #: ab3576, Lot: GR208064-1), and (C) GPER (anti-GPER antibodies, HPA027052, Sigma-Aldrich, Cat. #: HPA027052, Lot: A61748) IHC from adjacent control endometrial tissue (Control) and EC tissue (Tumor).
Fig 2: ERß mRNA and protein levels in endometrial cancer and adjacent control endometrium. (A) Before-and-after graph shows the normalized expression levels of the ESR2 gene in control endometrial tissue (Control) and the corresponding EC tissue (Tumor). The levels of gene expression are on a logarithmic scale. (B) ECL detection of ERß (59 kDa band). 18 paired samples were analyzed using anti-ERß antibodies (ab3576, Abcam, Cat. #: ab3576, Lot: GR208064-1) and GAPDH was used as a normalization control. Before-and-after plots show quantification of Western blotting data. Below, representative membrane with ERß and GAPDH staining is shown. EC tissue (T), adjacent control endometrial tissue (C), placenta (P) was used as a control tissue. (C) IHC scores in 21 samples from adjacent control endometrial tissue (Control) and EC tissue (Tumor). Tables show mean scores ± standard deviations, while before-and-after graphs show nuclear and cytoplasmic ERß (anti-ERß antibodies, 14C8, GeneTex, Cat. #: GTX70174, Lot: 20882 (1:100). (D) IHC staining in representative paired adjacent control endometrial tissue (C38) and EC tissue (T38) for ERß. In the negative controls (Control), the primary antibodies were replaced with serum of the same animal species (rabbit, mouse). Anti-ERß antibodies were validated (Supplementary Figure S6 and Table S5).
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