PGR siRNA reduced the expression of PTG synthases and
transporters and PGR binds to promoter regions of
PTGS2, PTGES, and
SLCO2A1. (A) Primary hGLCs were treated with or
without hCG (1 IU/mL) in the presence of negative control (NC) siRNA and
PGR siRNA (#1 and #2) for 10 hours. The levels of
mRNA for PTG synthases (PTGS2, PTGES,
and AKR1C1), transporters (SLCO2A1 and
ABCC4) and PGR were measured by
quantitative PCR. The levels of transcripts were normalized to
RNA18S5 mRNA in each sample (n = 3 independent
experiments). Bars with no common superscripts in each time point are
significantly different (P < 0.05). (B) ChIP
assay was performed using hGLC treated with hCG (1 IU/mL) for 12 hours.
DNAs immunoprecipitated with PGR antibody or IgG and input DNA were
analyzed using primer pairs specific to promoter regions of the
PTGS2, PTGES, and
SLCO2A1 gene and represented as arrows. Amplified
DNA fragments containing PGR responsive elements are represented as a
black box with the indicated PCR product size. Experiments were repeated
three times, each with independent cultured cells.