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. Author manuscript; available in PMC: 2015 Nov 1.
Published in final edited form as: Mol Immunol. 2014 Jun 25;62(1):54–62. doi: 10.1016/j.molimm.2014.05.019

Figure 4. NF-κB subunit p65, but not p50, is required for IL-1β-dependent activation of the TNFα gene.

Figure 4

A. 832/13 cells were transduced with adenoviruses overexpressing βGalactosidase (βGAL) or IκBα Superrepressor (SR). Following a 24 h transduction with the indicated adenoviruses, cells were stimulated for 15 or 30 min with 1 ng/mL IL-1β. An immunoblot was performed using whole cell lysates and antibodies against IκBα using actin as the loading control. B. 832/13 cells were transduced with adenoviruses overexpressing βGAL or IκBα SR. Following a 24 h exposure to adenoviruses, cells were stimulated for 3 h with 1 ng/mL IL-1β. **, P < 0.01. C, D. . 832/13 cells were transfected with siRNA targeted to p65 and incubated for 48 h. C. 48 h post-transfection, whole cell lysates were blotted to determine abundance of p65. Actin served as the loading control. D. After 48 h exposure to siRNA cells were stimulated for 3 h with 1 ng/mL IL-1β. ***, P < 0.001, n.s. = not significant. B-D. mRNA levels of TNFα were measured and normalized to RS9. Immunoblots were performed on two separate occasions and a representative image is shown. Data are shown as means ± SEM from three independent experiments.