Figure 4

RT-PCR reactions for LGR7 gene transcripts in human uterine cells. A. Primary cultures of endometrium stromal cells (ESC), glandular epithelial cells (EPI) and myometrial smooth muscle cells (MYO) cultivated for 6 days and treated or not, as indicated with 2.5 × 10⁷M medroxyprogesterone acetate (MPA) and/or 100 ng/ml porcine relaxin. The LGR7 PCR used primers #174 and #178, that for GAPD used #150 and #151. B. Primary cultures of endometrial stromal cells cultured for 3 or 6 days as indicated with progesterone (prog., 2.5 × 10^-7M), relaxin (100 ng/ml), rolipram (ro., 100 μM) and their combinations. RT-PCR products for prolactin (PRL), IGF-BP1 and GAPD mRNAs were performed in parallel on 6 day cultures only. All lanes are from the same gels run and photographed under identical conditions; the images have been cut to eliminate unwanted tracks only. PCR primers for LGR7 were as above, for GAPD #400 and #401 were used. C. RT-PCR reactions for RNA from a single preparation of ESC cells using different primer conditions (see lower panel) in order to determine the expression of possible LGR7 splice variants. The sizes of the DNA markers are indicated on the left of the upper panel. In the lower panel the sizes of the expected products, based on the published cDNA sequence, are indicated above the horizontal arrows. Primer #180 is positioned to include the start codon of the open reading frame, whereas #189 and #190 are both within the 5' untranslated region.