Figure 9. Role of DNA binding and protein–protein interactions on regulation of G6Pase promoter activity by NFκB.
(A) Western Blot analysis of HEK293 cells transiently transfected with 10 μg of empty vector (lane 1), wild-type pRSV-NFκB (lane 2), pRSV-NFκB-S276A (lane 3) or pRSV-NFκB-Y36F (lane 4). Western blotting was performed using an antibody against the p65/RelA subunit of NFκB. (B, C) H4IIE cells were co-transfected with 8.5 μg/dish of the reporter gene plasmids G6Pase(−1227/+57) (B) or pGL-TK-2×NFκB (C) together with 0.5 μg/dish of pRL control plasmid and 2 μg/dish of pRSV-NFκB, pRSV-NFκB-Y36F, pRSV-NFκB-S276A or pRSV-βgal in the presence or absence of TNFα as indicated. Data are presented as means±S.E.M. (n=3) of promoter activity compared with the respective luciferase activity in cells transfected with pRSV-βgal without TNFα, which was set as 100; *P<0.05 compared with expression in the presence of pRSV-βgal and in the absence of TNFα; **P<0.05 compared with expression in the presence of pRSV-βgal and pRSV-NFκB.