Figure 4.

The effect of FCF analogs on the expression of HE4. (A) HE4 overexpression clones (OVCAR8-C5) were treated with or without FCF (300 µM) for 7 h, after which the cell lysates and matching culture media were collected and analyzed for the levels of HE4 using enzyme immunoassays. The amount of secreted HE4 was normalized to the protein concentration of respective cell lysate. (B) ECC-1 and HCH-1 cells were treated with the indicated concentrations of FCF for 5 h and the secreted levels of HE4 were measured. (C) ECC-1 cells were treated with either UR214-1, UR214-7 or FCF at the concentrations indicated for 5 h at which point the secreted levels of HE4 were determined. (D) ECC-1 cells were washed and incubated with FCF (300 µM) for 5 h in basal media. Relative HE4 expression was determined by real time PCR (normalized to B2M). (E) ECC-1 cells were transfected with septin-2 targeting siRNA or non-targeting control siRNA for 48 h (bottom) or for 72 h (top). Relative gene expression of HE4 was determined by real time PCR (normalized to TBP; bottom). The intracellular levels of HE4 were measured using enzyme immunoassays (top).