Conditioned medium induces expression of COX-2 and prostaglandin F2α secretion. Expression of (A) COX-1 and (B) COX-2 mRNA in HUVECs incubated with V CM or P CM for 1,2,3,4,6,16 and 24 hrs as determined by quantitative RT-PCR analysis. (C) COX-2 mRNA in HUVECs incubated with V CM or P CM for 3 hrs in the absence/presence of FGFR1 tyrosine kinase inhibitor SU4984 or ERK1/2 inhibitor PD98059 as determined by quantitative RT-PCR analysis. Data are expressed as fold increase over control V CM (where V CM = 1, not shown). HUVECs were incubated with V CM or P CM for 0, 3, 6 and 16 hrs and thereafter conditioned medium from HUVECs was collected and used for PGE2 (D) and PGF2α (E) ELISA analysis as described in the methods. (F) HUVECs were treated with V CM, P CM or P CM and NS398 or indomethacin (Indometh.) for 6 hrs after which the concentration of PGF2α in the HUVEC medium was determined by ELISA. Data are represented as mean ± SEM. (* represents statistical significance P < 0.05).
Keightley et al. BMC Cell Biology 2010 11:8 doi:10.1186/1471-2121-11-8