Journal: Oncoimmunology

Article Title: Autoantibodies specific to estrogen receptor alpha act as estrogen agonists and their levels correlate with breast cancer cell proliferation

doi: 10.1080/2162402X.2015.1074375

Figure Lengend Snippet: Anti-ERα Abs reacted to binding domain of mERα on MCF-7 breast cancer cells. (A) Amino acid sequence of ERα (UniProtKB/Swiss-Prot: P03372.2). aPEPperMAP® Epitope Mapping (PEPperPRINT) defined the main reactive epitope (underlined) into the ligand binding domain (bold). (B) Representative flow cytometric analysis of mERα in MCF-7 cells by anti-ERα C-542 mAb (left panels) and human purified anti-ERα Abs (right panels) in untreated- (upper panels) or E2-BSA-treated (bottom panels) cells. Isotype control staining is represented by the broken line and anti-ERα-labeled cells are represented by the solid line. MFI, mean fluorescence intensity. A representative experiment out of five is shown. (C) Values of mERα/isotype mean fluorescence intensity ratio are reported (rMFI). Data are expressed as mean ± SD of five independent experiments. (D) Immunofluorescence analysis of mERα in MCF-7 cells by anti-ERα C-542 mAb (left panels) and human purified anti-ERα Abs (right panels) in untreated- (upper panels) or E2-BSA-treated (bottom panels) cells.

Article Snippet: Anti-ERα Abs reacted to binding domain of mERα on MCF-7 breast cancer cells. (A) Amino acid sequence of ERα (UniProtKB/Swiss-Prot: P03372.2). aPEPperMAP® Epitope Mapping (PEPperPRINT) defined the main reactive epitope (underlined) into the ligand binding domain (bold). (B) Representative flow cytometric analysis of mERα in MCF-7 cells by anti-ERα C-542 mAb (left panels) and human purified anti-ERα Abs (right panels) in untreated- (upper panels) or E2-BSA-treated (bottom panels) cells.

Techniques: Binding Assay, Sequencing, Ligand Binding Assay, Purification, Control, Staining, Labeling, Fluorescence, Immunofluorescence

Journal: Journal of cancer metastasis and treatment

Article Title: Osteolytic effects of tumoral estrogen signaling in an estrogen receptor-positive breast cancer bone metastasis model

doi: 10.20517/2394-4722.2021.27

Figure Lengend Snippet: Comparison of osteolytic ER+ BMET progression in young vs. skeletally mature mice supplemented with 0.72 mg E 2 . (A) Osteolytic lesion incidence and (B) osteolytic lesion area in young (5-week-old) and skeletally mature (16-week-old) mice supplemented with 0.72 mg E 2 and inoculated with ER+ tumor cells ( n = 8–13/group). Inset, representative immunohistochemical (IHC) images demonstrating cytokeratin+ (left panel; brown), ERα+ (right panel; brown) human breast cancer tumors in tibiae. *** P ≤ 0.001, **** P ≤ 0.0001 young vs. skeletally mature mice, by 2-way ANOVA with Sidak’s post-test. There was no significant difference (n.s.) in osteolytic lesion incidence by Log-rank (Mantel-Cox) test. (C) Representative hind limb radiographs in young (top) vs. mature (bottom) age-matched control (left panels), naive E 2 (0.72 mg)-supplemented (middle panels), or tumor cell-inoculated and E 2 (0.72 mg)-supplemented mice (right panels) 6 weeks post-inoculation and E 2 supplementation. Osteolytic lesions are marked by arrows.

Article Snippet: For measuring histologic breast cancer tumor size (tumor burden), epithelial MCF-7 breast cancer tumors were identified using a pan-cytokeratin primary antibody (#Z0622, Agilent Dako, Santa Clara, CA) and continued expression of ERα was verified using a human ERα primary antibody (#ab108398, Abcam, Cambridge, United Kingdom) using previously described immunohistochemical (IHC) methods [ ] .

Techniques: Immunohistochemical staining

Journal: Toxicological Sciences

Article Title: Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase

doi: 10.1093/toxsci/kfz004

Figure Lengend Snippet: Comparison of the HEK293 ERE/Gal4-Lux Autobioluminescent Assay and the HeLa-9903 Estrogen Receptor Transcription Activation Assay

Article Snippet: HEK293 ERE/Gal4-Lux and MCF7 ERE/Gal4-Lux molecular assembly Human ERα cDNA was synthesized (GenScript) and cloned into the pcDNA3.1/Zeo vector (Life Technologies).

Techniques: Comparison, Activation Assay

Journal: Toxicological Sciences

Article Title: Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase

doi: 10.1093/toxsci/kfz004

Figure Lengend Snippet: Comparison of the HEK293 ERE/Gal4-Lux Autobioluminescent Assay and the H295R Steroidogenesis Assay

Article Snippet: HEK293 ERE/Gal4-Lux and MCF7 ERE/Gal4-Lux molecular assembly Human ERα cDNA was synthesized (GenScript) and cloned into the pcDNA3.1/Zeo vector (Life Technologies).

Techniques: Comparison

Journal: Toxicological Sciences

Article Title: Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase

doi: 10.1093/toxsci/kfz004

Figure Lengend Snippet: EC 50 Values of Common Endocrine Disruptor Chemicals Tested Using HEK293 ERE/Gal4-Lux Autobioluminescent Cells

Article Snippet: HEK293 ERE/Gal4-Lux and MCF7 ERE/Gal4-Lux molecular assembly Human ERα cDNA was synthesized (GenScript) and cloned into the pcDNA3.1/Zeo vector (Life Technologies).

Techniques:

Journal: Toxicological Sciences

Article Title: Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase

doi: 10.1093/toxsci/kfz004

Figure Lengend Snippet: Qualitative ER Agonism Responses of the HEK293 ERE/Gal4-Lux Autobioluminescent Assay Relative to the Multi-Assay ICCVAM Meta-Analysis for All Tested Compounds

Article Snippet: HEK293 ERE/Gal4-Lux and MCF7 ERE/Gal4-Lux molecular assembly Human ERα cDNA was synthesized (GenScript) and cloned into the pcDNA3.1/Zeo vector (Life Technologies).

Techniques: