Figure 3

EGF- or TNF-responsive chemokine signatures in CaOV-3 cells. EGF- or TNF-responsive chemokines (A) and chemokine receptors (B) were determined by PCR array. Cells were incubated with vehicle (Control), EGF (10 ng/ml), TNF (10 ng/ml) or both for 1 h. Values shown are fold changes compared to vehicle controls. After isolating total RNA, a real-time PCR was performed using a customized human chemokine PCR array. Dashed lines indicate the equivalent of 2-fold changes from controls. A chemokine signature with a >2-fold increase was considered as EGF- or TNF-responsive. White, blue, green and red blocks indicate an absent, low, transient (low to high) and high expression level of chemokines, respectively, based on a >35, 30–35, 30–35 to <30 and <30, on average, threshold cycle. An asterisk (*) indicates EGF- or TNF-responsive chemokines with a high expression level (below an average threshold cycle of 30). (C) Effects of EGF or TNF (or both) on IκB, Akt and Erk activation. Cells were treated with EGF (10 ng/ml) or TNF (10 ng/ml) for 0–60 min. Whole cell lysates were prepared and IκB, Akt and Erk activations were confirmed by Western blot analyses. Experiments were performed in duplicate and a representative result is shown.