Figure 3.

GLP2 Inhibits the Expression of NF-κB in MG63

(A) ChIP assay with anti-RNA polII followed by PCR with promoter DNA primers of NF-κB in MG63 cells infected with rLV-GLP2 or rLV. IgG ChIP was the negative control. NF-κB promoter as INPUT. (B) The assay of luciferase activity of prmoter of NF-κB in MG63 infected with rLV-GLP2 or rLV, respectively. Data are means of value from three independent experiment (error bar ± SEM; **p < 0.01). (C) Western blotting with anti-NF-κB and RT-PCR with NF-κB primers in MG63 infected with rLV-GLP2 or rLV. β-actin as internal control. (D) Co-immunoprecipitation (coIP) with anti-NF-κB followed by western blotting with anti-β-catenin and anti-AP1 in MG63 infected with rLV-GLP2 or rLV, respectively. IgG IP was used as a negative control. INPUT refers to western blotting with anti-β-catenin and anti-AP1. (E) ChIP assay with anti-NF-κB followed by PCR with DNA primers (promoter region) of c-Myc,PKM2, CyclinD1 in MG63 cells infected with rLV-GLP2 or rLV. IgG ChIP was the negative control. The promoters of c-Myc, PKM2, CyclinD1 as INPUT. (F) RT-PCR with primers of c-Myc, PKM2, CyclinD1 in MG63 infected with rLV-GLP2 or rLV. β-actin as internal control. (G) Western blotting with anti-c-Myc, anti-PKM2, anti-CyclinD1 in MG63 infected with rLV-GLP2 or rLV. β-actin as internal control.