Figure 4.

Excessive NF-κB Abrogates the Function of GLP2 in MG63 In Vitro and In Vivo

(A) Western blotting with anti-GLP2, NF-κB in MG63 infected with rLV-GLP2, rLV-GLP2 plus pcDNA3-NF-κB or pLV. β-actin was used as an internal control. (B) Cell proliferation assay was performed in 96-well format using the CCK8 cells proliferation kit to determine the cell viability as described by the manufacturer. Each sample was assayed in triplicates for 3 days consecutively. Cell growth curve was based on the corresponding the relative values of OD450 and each point represents the mean of three independent samples. Data are means of value from three independent experiments (error bar ± SEM; **p < 0.01 and *p < 0.05). (C) Cell BrdU assay. Data are means of value from three independent experiment (error bar ± SEM; **p < 0.01). (D) Cell plate colony formation ability assay. Data are means of value from three independent experiment (error bar ± SEM; **p < 0.01). (E) The xenograft tumors from BALB/c nude mouse injected with MG63 cells infected with rLV, rLV-GLP2, rLV-GLP2 plus pcDNA3-NF-κB subcutaneously at armpit. The xenograft tumors weight (gram) in the three groups. Data were means of value from six BALB/c mice (mean ± SEM; n = 6; **p < 0.01). (F) A portion of each xenograft tumor was fixed in 4% formaldehyde and embedded in paraffin, and the micrometers of sections (4 μm) were made for PCNA staining (original magnification × 100). **p < 0.01.