gper agonist g1  (Tocris)

Journal: Cancers

Article Title: G Protein-Coupled Oestrogen Receptor Actions Targeting the Hallmarks of Cancer in Human Prostate Cells: From Cell Fate to Metabolic Reprogramming

doi: 10.3390/cancers18071137

Figure Lengend Snippet: Representative images of GPER high vs. low immunoreactivity in human benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) lesions and prostate adenocarcinoma. Haematoxylin–eosin counterstaining magnification 40×. Scale bar 200 μm for BPH and 100 μm for PIN and adenocarcinoma.

Article Snippet: Subsequently, cells were treated with increasing concentrations of the GPER agonist G1 (Tocris Bioscience, Madrid, Spain), or vehicle for 24 h. The tested concentration range, from 0.001 to 100 μM, was selected based on previous in vitro studies investigating GPER activation, which showed that micromolar concentrations are commonly required to elicit measurable anti-proliferative effects [ ].

Techniques:

Journal: Cancers

Article Title: G Protein-Coupled Oestrogen Receptor Actions Targeting the Hallmarks of Cancer in Human Prostate Cells: From Cell Fate to Metabolic Reprogramming

doi: 10.3390/cancers18071137

Figure Lengend Snippet: GPER expression in non-neoplastic PNT1A and neoplastic LNCaP, DU145 and PC3 human prostate cells. ( A ) Protein expression determined by WB analysis after normalisation with α-tubulin. Error bars indicate mean ± S.E.M. (n = 3) *** p < 0.001. ( B ) Representative immunoblots. ( C ) Representative images of GPER subcellular localisation obtained with a Zeiss LSM 710 laser scanning confocal microscope under 400× magnification. Nuclei are stained blue and GPER green. Original western blots are presented in .

Article Snippet: Subsequently, cells were treated with increasing concentrations of the GPER agonist G1 (Tocris Bioscience, Madrid, Spain), or vehicle for 24 h. The tested concentration range, from 0.001 to 100 μM, was selected based on previous in vitro studies investigating GPER activation, which showed that micromolar concentrations are commonly required to elicit measurable anti-proliferative effects [ ].

Techniques: Expressing, Western Blot, Microscopy, Staining

Journal: Cancers

Article Title: G Protein-Coupled Oestrogen Receptor Actions Targeting the Hallmarks of Cancer in Human Prostate Cells: From Cell Fate to Metabolic Reprogramming

doi: 10.3390/cancers18071137

Figure Lengend Snippet: GPER subcellular localisation in non-neoplastic PNT1A and neoplastic LNCaP, DU145 and PC3 human prostate cells. ( A ) Pearson’s correlation coefficient for the GPER co-localisation with the cell membrane and endoplasmic reticulum. ( B ) Pearson’s correlation coefficient for the GPER co-localisation with the cell membrane and endoplasmic reticulum after GPER activation by G1 (1 µM). Results are expressed as fold-change relative to the control untreated group (0 μM G1, dashed line). Error bars indicate mean ± S.E.M. (n = 3) * p < 0.05 ** p < 0.01 *** p < 0.001. ( C , D ) Representative confocal microscopy images showing the co-localisation of GPER (green) with cell membrane (WGA in red), endoplasmic reticulum (calnexin in red) or nucleus (Hoechst in blue) in control and G1-treated (1 µM) cells. Images were obtained in the Zeiss LSM 710 laser scanning confocal microscope under 400× magnification. Negative controls obtained by omission of the primary antibody or WGA are provided as insert panels (−).

Article Snippet: Subsequently, cells were treated with increasing concentrations of the GPER agonist G1 (Tocris Bioscience, Madrid, Spain), or vehicle for 24 h. The tested concentration range, from 0.001 to 100 μM, was selected based on previous in vitro studies investigating GPER activation, which showed that micromolar concentrations are commonly required to elicit measurable anti-proliferative effects [ ].

Techniques: Membrane, Activation Assay, Control, Confocal Microscopy, Microscopy

Journal: Cancers

Article Title: G Protein-Coupled Oestrogen Receptor Actions Targeting the Hallmarks of Cancer in Human Prostate Cells: From Cell Fate to Metabolic Reprogramming

doi: 10.3390/cancers18071137

Figure Lengend Snippet: Effect of androgens in regulating GPER expression in LNCaP, DU145 and PC3 human PCa cells. ( A ) GPER protein expression after treatment with DHT (10 nM) for 24 or 48 h. Protein expression was determined by WB analysis after normalisation with β-actin. Results are expressed as fold-change relative to the control untreated group (0 nM DHT, dashed line). Error bars indicate mean ± S.E.M. (n = 5) * p < 0.05. ( B ) Representative immunoblots. Original western blots are presented in .

Article Snippet: Subsequently, cells were treated with increasing concentrations of the GPER agonist G1 (Tocris Bioscience, Madrid, Spain), or vehicle for 24 h. The tested concentration range, from 0.001 to 100 μM, was selected based on previous in vitro studies investigating GPER activation, which showed that micromolar concentrations are commonly required to elicit measurable anti-proliferative effects [ ].

Techniques: Expressing, Control, Western Blot