Fig. 2.
NF-κB positively regulates TGF-α expression. (A) Astrocytes were co-transfected with 0.5 μg of wild type (WT) or triple mutant (TM) and 0.1 μg of either empty vector (EV) prcRSV or p65, followed by the measurement of TGF-α promoter activity. (B) Cells were transfected overnight with 0.5 μg of WT, single (−53, −218 and −507 position), double (−53/−507, −53/−218 and −218/−507 positions) or triple NF-κB mutants (all three mutant NF-κB sites in −53, −218 and −507 positions), followed by the luciferase assay to determine TGF-α promoter activity. (C) Cells were transfected overnight with 2 μg of EV prcRSV or p65 followed by TGF-α mRNA analysis by qPCR. The overexpression of p65 was confirmed by western blotting. (D) Cells were transfected overnight with 2 μg of EV pCMV or IκBα, followed by TGF-α mRNA analysis by qPCR. The overexpression of IκBα was confirmed by western blotting. (*p<0.05, ***p<0.001, #p<0.05, ###p<0.001; significantly different from the control; N.S., not significant; Student’s t-test, one-way and two-way ANOVA followed by Tukey’s post hoc test; N=3).
