Effects of IKK inhibition on nuclear translocation of NFκB p50 and
p65 in Grx1 overexpressing rMC-1 cells. rMC-1 cells were cultured in
normal glucose (5 mm) medium, pretreated with Bay 11-7085
(Bay) for 30 min prior to Grx1 overexpression, immediately infected
with m.o.i. of 10 of either Ad-Grx1 or Ad-Empty, cultured for two more days in
normal glucose medium, and collected. Cells were separated into nuclear
fractions, run on a 12% SDS-PAGE, and immunoprobed for p50 (1:1,000), p65
(1:3,000), and the nuclear loading control YY1 (1:1,000). Ad-Grx1 increased
p50 in the nucleus by 4-fold (±0.7) (n = 9) (panel
a), *, p ≤ 0.002. Cells overexpressing Grx1 treated with 10
and 20 μm Bay 11-7085 were significantly decreased from
Grx1-overexpressing cells with no inhibitor (n = 9) (panel
a), #, p < 0.02. Ad-Grx1 increased p65 in the nucleus by
nearly 2-fold (±0.7) (n = 11) (panel b), *,
p ≤ 0.002. Cells overexpressing Grx1 treated with 10 and 20
μm Bay 11-7085 were significantly decreased from
Grx1-overexpressing cells with no inhibitor (n = 11) (panel
b), #, p < 0.02.