Figure 4

EGF- or TNF-responsive chemokine signatures in TOV-21G 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 hour. 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. Dotted lines indicate the equivalent of 2-fold changes from controls; chemokine signature with a greater than 2-fold increase is recognized as EGF- or TNF-responsiveness. 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. 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) for 0 to 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.