Fig. 4

NF-κB activation in HPMCs. a Total and phosphorylated NF-κB p65 expression and localization were determined in two HPMCs samples (Meso 4 and Meso 7 cells) using anti-NF-κB p65 specific antibodies. HPMCs were incubated with either FBS (10%), TNF-α (20 ng/ml), OVC346 (10%) or OVC439 (10%) for 20 min. b Meso7 cells were incubated with OVC508 ascites (10% v/v) in the presence or absence of NFκB inhibitor BAY117082 (5 μM) for 24 h. Cells were then washed three times with PBS and incubated with standard medium for 24 h. Lysates were then obtained and immunoblotted for MUC16, c-Src and phosphorylated (Y418) c-Src. GAPDH was used as a loading control. c Meso 7 cells were pretreated or not with BAY117082 (2 or 5 μM) for 1 h prior to the addition of ascites OVC508 or OVC690 (10%) to the medium. Cells were incubated overnight. The next day, cells were washed 3 times and incubated 24 h with standard medium. The medium was removed and MUC16 concentration were measured in the condition medium. Results are from two independent experiments performed in duplicates