Time course of C/EBPβ, phospho-IκB (p-IκB), IκB, and p65 proteins in response to cAMP and TNF. Panel A, Effects of cAMP and TNF on the time course of C/EBPβ, p-IκB, IκB, and p65 proteins in mouse granulosa cells. Cells were incubated with TNF (10 ng/ml) alone, cAMP (500 μm) alone, and in combination for 3, 6, 12, and 24 h. Each gel lane was loaded with 20 μg of total protein using whole-cell lysates. β-Actin was used as a loading control. Panel B, Nuclear localization of C/EBPβ protein by immunocytochemistry after cAMP and/or TNF in mouse granulosa cells. The experiments were repeated twice and a representative figure is shown. C, Control; A, cAMP; T, TNF; TA, TNF+cAMP.