Figure 4:
Exogenously overexpressed GBP-5 abrogates IL-1β-induced chemokine production and adds to the protective action of IFN-γ in human RASFs. Human RASFs were transduced with control (NC) and GBP-5 lentiviral particles at 3MOI for overnight. After 48 h, cells were treated with IL-1β or IFN-γ for 24 h. Whole Cell extracts were collected to determine the expression of GBP-5. A, Western immunoblotting showing the exogenous expression of GBP-5 overexpression. B, The ELISA result showed significant reduction in IL-1β-induced IL-6, IL-8, and ENA-78/CXCL5 production. C, Human RASFs were transduced as described in (A) and treated with IL-1β for 30 min. Western immunoblotting analysis showed a marked decreased in the expression of IL-1β-induced level of p-c-Jun, p-NF-κBp65, p-JNK, p-ERK1/2, and p-P38. β-actin was tested as a loading control. D-F, Human RASFs were transfected with empty vector or GBP-5 plasmid (1μg/ml) in 12-well plates for 48 h. Cells were treated with IL-1β (10 ng/ml) or IL-1β and IFN-γ at the indicated concentration for 24 h. Conditioned media was collected to determine the level of IL-6, and IL-8, and ENA-78/CXCL5 production. *p<0.05 or **p<0.01 vs NC; #p<0.05 or ##p<0.01 vs NS; *p<0.05 or **p<0.01 vs IL-1β.