Journal: Pharmacological Research

Article Title: The flavonoid agathisflavone modulates the microglial neuroinflammatory response and enhances remyelination

doi: 10.1016/j.phrs.2020.104997

Figure Lengend Snippet: Estrogen receptor (ER) activation is required for agathisflavone to inhibit microgliosis and promote remyelination. (A) Root mean square deviation (RMSD) values and rod representation of crystallographic ligand pose (lilac) and the best pose of this ligand generated by DOCK 6.8 (yellow) for each complex. Distances less than 2 Å between the calculated pose and the crystallographic pose indicates that the program was successful in reproducing the experimental data (B) Representation of interactions between agathisflavone (FAB) and retinoic and estrogen receptors; the captions are described in the figure. (C–G) Organotypic cerebellar slices from SOX10-EGFP animals were maintained for 7DIV, then exposed to LPC for 15–17 h, followed by 2 h pretreatment with the selective ER-α antagonist MPP dihydrochloride at 10 nM (1,3-Bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole-dihydrochloride), or the selective ER-β antagonist PHTPP at 1 μM (4-[2-Phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5- a ]pyrimidin-3-yl]phenol), which were kept together with 10 μM FAB for a further 2 DIV. (C) Oligodendrocytes were identified by the Sox10-EGFP reporter (green) and slices were immunolabeled for MBP (red), neurofilament (blue) and Iba-1 (yellow); scale bar: 20 μm. Bar graphs compare LPC and LPC + FAB 10 μM with the effects of the ER antagonists MPP and PHTPP on the NF + axon index (D), the percentage of MBP+/NF + myelinated axons (E), the number of Sox10-EGFP + oligodendrocytes (F) and the number of Iba1+ microglia (G); data are expressed as the mean ± SEM (n = 5); ‡ p < 0.05, ‡‡ p < 0.01, ‡‡‡‡ p < 0.0001 (comparing LPC-DMSO to other treatment groups); †† p < 0.01 and ††† p <0.001 (comparing LPC + FAB10 to LPC + FAB10+MPP); & p < 0.05 (comparing LPC+FAB10+MPP to LPC+FAB10+PHTPP); One-way ANOVA followed by Tukey’s post-hoc test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: To assess the potential involvement of estrogen receptors (ER) on the effects of agathisflavone following LPC treatment, slices were pre-incubated for 2 h in medium containing the selective ER-α antagonist MPP dihydrochloride at 10 nM (1,3-Bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride; Sigma), or the selective ER-β antagonist PHTPP at 1 μM (4-[2-Phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5- a ]pyrimidin-3-yl]phenol; Tocris).

Techniques: Activation Assay, Generated, Immunolabeling

Journal: British Journal of Cancer

Article Title: The DNA mismatch repair gene hMSH2 is a potent coactivator of oestrogen receptor α

doi: 10.1038/sj.bjc.6602614

Figure Lengend Snippet: In vivo association between hMSH2 and ER α / β and in vitro association between hMSH2 and ER α / β. ( A ) (i) Ishikawa cells were lysed and subjected to immunoprecipitation (IP) with the antbodies to ER α and IgG. MDA-MB-231 cell lysate was immunoprecipitated with the antbodies to ER β BRCA1 and IgG. The immunoprecipitates were separated by SDS–PAGE and analysed by immunoblotting (IB) with the anti-hMSH2 antibody. (ii) Reciprocal IP was performed to detect endogenous hMSH2-ER α and hMSH2-ER β interactions by IB. ( B ) In vitro translated 35 S-labelled hMSH2 was pulled down by GST-ER α / β AF-1 or GST-ER α / β AF-2. At the same time, in vitro translated TRAP220 was incubated with GST-ER α AF-2. The mixtures were washed and subjected to SDS–PAGE and analysed.

Article Snippet: Oestrogen receptor α / β -positive T-47D (HTB-133) and ER β -positive MDA-MB-231 (HTB-26) human breast cancer cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, In Vitro, Immunoprecipitation, SDS Page, Western Blot, Incubation

Journal: International Journal of Endocrinology

Article Title: Estrogen Induces Metastatic Potential of Papillary Thyroid Cancer Cells through Estrogen Receptor α and β

doi: 10.1155/2013/941568

Figure Lengend Snippet: E2 induces metastasis via downregulation of E-cadherin and upregulation of vimentin and MMP-9. BCPAP cells were treated with vehicle (DMSO) alone, 10 −8 M E2, 10 −6 M PPT, or 10 −6 M DPN. Whole cell lysates were extracted, and E-cadherin, vimentin, and MMP-9 protein were detected by western blot analysis. β -actin was used as a loading control. * P < 0.05 compared with control.

Article Snippet: Cells were then incubated in fresh medium with vehicle (DMSO) alone, 10 −8 M E2 (Sigma, St. Louis, MO), 10 −6 M PPT (an ER α -selective agonist) (Tocris, Ballwin, MO), or 10 −6 M DPN (an ER β -selective agonist) (Tocris, Ballwin, MO) for 24 h. Concentrations of E2, PPT, and DPN are used according to the report by Zeng et al. Control cultures received the same volume of DMSO.

Techniques: Western Blot, Control

Journal: Frontiers in Oncology

Article Title: Andrographolide Inhibits ER-Positive Breast Cancer Growth and Enhances Fulvestrant Efficacy via ROS-FOXM1-ER-α Axis

doi: 10.3389/fonc.2022.899402

Figure Lengend Snippet: AD inhibits ESR1 transcription in ER-positive breast cancer. (A) Volcano plot showing the DEGs between the control and AD treatment group from the GSE85871 dataset. (B) KEGG pathway enrichment analysis for the DEGs and qvalue refers to -log 10 (pvalue). (C) Venn plot revealing the overlapping target genes for AD against ER-positive breast cancer. Blue circle represents AD; red circle represents ER-positive breast cancer. (D) Network of target genes for AD against ER-positive breast cancer was built using the STRING database and Cytoscape software. Blue diamond represents AD; red hexagon represents ER-positive breast cancer; blue circles represent target genes of AD in ER-positive breast cancer and were arranged from the inner circle to the outer circle according to the degree of association. (E) Cells were treated with 1‰ DMSO or the indicated concentrations of AD for 24 h. Cell protein was extracted and detected by Western blotting with antibodies against ER-α, ER-β and β-actin. (F) The mRNA level of ESR1 was quantified by RT-PCR (normalized to β-actin). (G) Cells were treated with AD (40 μM) for 0 h, 6 h, 12 h, 24 h, respectively. Cell protein was extracted and detected by Western blotting with antibodies against ER-α, ER-β and β-actin. (H) The mRNA level of ESR1 was quantified by RT-PCR (normalized to β-actin). (I) Cells were treated with 1‰ DMSO or the indicated concentrations of AD for 24 h. Cell protein was extracted and detected by Western blotting with antibodies against PR, C-MYC, Cathepsin D and β-actin. (J) Proteins extracted from tumor tissues of mice treated with 20% HPBCD or AD (150 mg/kg) were detected by Western blotting with antibodies against ER-α and β-actin. (Data were presented as mean ± SD. ** P < 0.01 and *** P < 0.001).

Article Snippet: After blocking with 5% nonfat milk in TBST, the membranes were incubated with the primary antibody as follows ER-α, FOXM1, Cathepsin D, β-tublin, LaminA/C (Cell Signaling Technology, Danvers, MA, USA), ER-β (AbSci, CA, USA), β-actin (HuaBio, Hangzhou, China), C-MYC and PR (Santa Cruz Biotechnology, Santa Cruz, CA, USA), then incubated with secondary antibody.

Techniques: Control, Software, Western Blot, Reverse Transcription Polymerase Chain Reaction