EGF- or TNF-responsive chemokine signatures in SKOV-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 in (A) and (B) indicate the equivalent of 2-fold changes from controls. A chemokine signature with a >2-fold increase was considered to be EGF- or TNF-responsive. White, blue, green and red blocks indicate absent, low, transient (low to high) and high expression levels 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 on IκB, Akt and Erk activation. Cells were treated with EGF (10 ng/ml) or TNF (10 ng/ml) to both 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.