Fig. 3

Ovarian cancer ascites and CCL18 induce the phosphorylation of Pyk2. a Serous ovarian cancer ascites OVC509 induce a rapid phosphorylation of Pyk2 at tyr402 in OVCAR3 and CaOV3 cells. Cells were incubated in serum-free media for 24 h, the media was change and OVC509 ascites (10 % v/v) was added for up to 90 min. Densitometric quantification of phosphorylated Pyk2 normalized to total Pyk2. The fold increase of phosphorylated Pyk2 is indicated at the bottom of pPyk2 panels. b The stimulation of Pyk2 phosphorylation was confirmed in CaOV3 and OVCAR3 cells using the additional serous ovarian cancer ascites, OVC439 and OVC551. c Immunoblot for total Pyk2 expression from CaOV3 and OVCAR3 lysates obtained at 24 h following the addition of ovarian cancer ascites. d Real-time PCR analysis of Pyk2 transcript levels at 2 h and 6 h following the addition of OVC439 ascites. Results were standardized using primers of the housekeeping gene RPLPO. Results are expressed as fold change relative to basal levels observed in cells incubated in the absence of ascites. e CaOV3 and OVCAR3 cells were treated with increasing concentration of CCL18 (0–50 ng/ml) or ascites for 90 min. Immunoblot were obtained to assess the phosphorylation of Pyk2