Figure 4.

Effect of FOS inhibitor on the production of progesterone and PGE₂ and on the expression of mRNA for PG synthases and transporters in hGLCs. Primary hGLCs were treated with or without T-5224 (FOS inhibitor, 20 µM) in the absence or presence of hCG (1 IU/mL). (A) The levels of PGE₂ and progesterone in media collected at 24 h were measured by PGE₂ (a) enzyme immunoassay and (b) progesterone assay, respectively. (B) The levels of mRNA for PG synthases (PTGS2, PTGES, and AKR1C1) and transporters (SLCO2A1 and ABCC4) at 12 h were measured by quantitative PCR. The levels of transcripts were normalized to those of RNA18S5 mRNA in each sample (n = 4 to 6 independent experiments). (C) (a) PTGES and SLCO2A1 were detected by Western blots. The membrane was reprobed with a monoclonal antibody against ACTB to assess the loading of protein in each lane. (b) The band intensities for each protein were measured by ImageJ and normalized to the intensity of β-actin (ACTB) in the corresponding sample. The experiments were repeated three times with an independent sample. Bars with no common superscripts in each time point are significantly different (P < 0.05). Con, control.