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Image Search Results
Journal: Cells
Article Title: miR-338-3p Is Regulated by Estrogens through GPER in Breast Cancer Cells and Cancer-Associated Fibroblasts (CAFs)
doi: 10.3390/cells7110203
Figure Lengend Snippet: c-Fos and miR-338-3p are involved in Cyclin D1 regulation in SkBr3 cancer cells and CAFs. Luciferase activity of Cyclin D1 reporter gene in SkBr3 cancer cells ( A ) and CAFs ( B ) transfected for 8 h with a vector or a dominant-negative c-Fos construct (DN-Fos) before treatment with 100 nM of E2 and 100 nM G-1 for 18 h. Luciferase activity of Cyclin D1 reporter gene in SkBr3 cancer cells ( C ) and CAFs ( D ) transfected for 24 h with 25 nM miR-Ctrl or miR-338-3p mimic (miR-338-3p m) in combination or not with 50 nM miR-338-3p inhibitor (miR-338-3p i) before treatment for 18 h with 100 nM E2 or 100 nM G-1. The luciferase activity was normalized to the internal transfection control, values of cells receiving vehicle (-) were set as 1-fold induction upon which the activity obtained upon the indicated treatments was calculated. mRNA expression of Cyclin D1 in SkBr3 cells ( E ) and CAFs ( F ) transfected for 48 h with 25 nM miR-Ctrl or miR-338-3p mimic (miR-338-3p m) in combination or not with 50 nM miR-338-3p inhibitor (miR-338-3p i) before treatment for 8 h with 100 nM E2 or 100 nM G-1. Each column represents the mean ± SD of three independent experiments performed in triplicate. (*) indicates p < 0.05 for cells receiving treatments vs cells treated with vehicle (-).
Article Snippet: Equal amounts of protein extract were resolved on SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, Italy), probed overnight at 4 °C with antibodies against: c-Fos (E-8, sc-166940) and β-Actin (AC-15, sc-69879) (Santa Cruz Biotechnology, DBA, Italy), GPER (AB137479) (Abcam, Euroclone, Milan, Italy) and
Techniques: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Dominant Negative Mutation, Construct, Expressing
Journal: Cells
Article Title: miR-338-3p Is Regulated by Estrogens through GPER in Breast Cancer Cells and Cancer-Associated Fibroblasts (CAFs)
doi: 10.3390/cells7110203
Figure Lengend Snippet: miR-338-3p prevents Cyclin D1 protein induction by E2 and G1 in SkBr3 cancer cells and CAFs. Cyclin D1 protein expression in SkBr3 cancer cells ( A , B ) and CAFs ( C , D ) transfected for 48 h with 25 nM miR-Ctrl or miR-338-3p mimic (miR-338-3p m) in combination or not with 50 nM miR-338-3p inhibitor (miR-338-3p i) before treatment for 12h with 100 nM E2 or 100 nM G-1. Side panels show densitometry analysis of the blots normalized to the loading control β-actin. (*) indicates p < 0.05 for cells receiving treatments vs cells treated with vehicle (-).
Article Snippet: Equal amounts of protein extract were resolved on SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, Italy), probed overnight at 4 °C with antibodies against: c-Fos (E-8, sc-166940) and β-Actin (AC-15, sc-69879) (Santa Cruz Biotechnology, DBA, Italy), GPER (AB137479) (Abcam, Euroclone, Milan, Italy) and
Techniques: Expressing, Transfection