Skip to main content
Fig. 5 | Molecular Cancer

Fig. 5

From: Epigenetic down regulation of G protein-coupled estrogen receptor (GPER) functions as a tumor suppressor in colorectal cancer

Fig. 5

Inhibition of NF-κB participated in G-1 induced growth arrest. HCT-116 cells treated with 1 μM G-1 for 0 to 60 min (a) or increasing concentrations of G-1 for 30 min (b), the total and phosphorylation of p65 were measured by western blot analysis; (c) HCT-116 cells were treated with or without G-1 for 6 h, and the subcellular localization of p65 (green) was examined by immunofluorescence staining and nuclei were stained with DAPI (blue); (d) HCT-116 cells were transfected with pGL3-Basic-luc reporter plasmid containing 5 copies of the κB site plasmid and pRL-TK plasmids which served as the correcting transfection efficiency and then treated with or without G-1 (1 μM) for 24 h, then the lysates were assayed. Shown are relative luciferase activities normalized to Renilla activities; (e) HCT-116 or SW480 cells were treated with 1 μM G-1 or 10 μM NF-κB inhibitor BAY11-7082 (BAY) for 48 h, the cell viability was measured by use of CCK-8 kit; (f) HCT-116 or SW480 cells were transfected with pcDNA3.1 (vector) or pcDNA3.1/p65 for 24 h and then treated with 1 μM G-1 for 48 h, the cell viability was measured by use of CCK-8 kit, *p < 0.05 compared with G-1 group.; (g, h, & i) HCT-116 cells were treated with 1 μM G-1 for the indicated times, proteins were examined by western blot analysis, mRNAs were measured by qRT-PCR; (j) HCT-116 cells were treated with G-1 for 12 h, and then p65 was immunoprecipitated from equal amount of lysates and the associated GSK-3β was detected by western blot analysis. Data were presented as means ± SD of three independent experiments. *p < 0.05, **p < 0.01 compared with the control group

Back to article page