Figure 2

Ligand-dependent/EGFR signaling triggers IL-6 production via NFkB activation. (a) IL-6 levels measured by ELISA on conditioned media from starved EOC cell lines left untreated, or treated for the indicated times with EGF (20 ng/ml) alone or together with AG1478 (20 μM). Representative growth curves of one of three independent experiments are shown. Each point represents the mean of five independent replicates ± s.d. (b) Real-time PCR for IL-6 from total RNA of starved EOC cell lines left untreated, treated for 24 h with EGF (20 ng/ml) alone or together with AG1478 (20 μM), LY294002 (50 μM) or with UO126 (40 μM). Results are presented as relative expression normalized to GAPDH mRNA levels. (c) Luciferase promoter gene assay of starved EOC cells transiently transfected with reporter plasmids containing the NFkB binding sites and stimulated for 24 h with EGF (20 ng/ml). (d) Inhibition of NFkB transcriptional activity of starved EOC cells transiently transfected as above and stimulated for 24 h with EGF alone or together with AG1478 (black), LY294002 (gray) and UO126 (white). (c, d) Data are mean values (±s.d.) normalized for transfection efficiency in a representative experiment.