05) and 247-fold (p<0.01), respectively (Figure 1A). The difference sellekchem in the PRLr mRNA synthesized between 2 and 4 h was significant (p<0.01) as well. The expression of PRL mRNA increased 80-fold (p<0.05) and 133-fold (p<0.01) after 1 and 2 h of LPS stimulation, respectively. Then, PRL mRNA decreased below 30-fold (p<0.01) at 4 h, increased 80-fold (p<0.05) at 8 h, and again decreased below 30-fold after 48 h (p< 0.05) (Figure 1B). Figure 1 PRLr mRNA and PRL mRNA synthesis in THP-1 monocytes activated with LPS using real-time PCR. THP-1 monocytes stimulated with LPS 1 ��g/ml were harvested at different times and three independent RT-PCR experiments were analyzed using comparative ... PRLr and PRL were expressed in THP-1 monocytes stimulated with LPS Two PRLr isoforms of 100 and 50 kDa were identified in THP-1 monocytes by Western blot until 8 h of stimulation with LPS (Figure 2A).

In relation to untreated cells, the expression of the 100 kDa isoform increased from 1 to 8 h, except at 4 h when the expression was significantly lower in comparison with any time after stimulation with LPS (p<0.01) (Figure 2A). A basal expression of the PRLr isoform of 50 kDa was observed (Figure 2A), but an increase in a time-dependent manner was found from 4 h (p<0.05) to 72 h (p<0.01) after LPS stimulation (Figure 2A). Also, a continuous increased expression of PRL of 60 kDa in THP-1 monocytes was detected from 1 to 8 h after stimulation with LPS; then, after 48 h the expression of a bigger PRL of 80 kDa was detected (Figure 2B). No basal expression of PRL was detected in untreated cells (Figure 2B).

Figure 2 PRLr and PRL expression in THP-1 monocytes activated with LPS using Western blot. THP-1 monocytes stimulated with LPS 1 ��g/ml were harvested at different times and two independent Western blot experiments were performed and analyzed using densitometry … Isoforms of PRLr and PRL were expressed by monocytes from healthy subjects after stimulation Cilengitide with LPS In untreated monocytes obtained from healthy subjects a PRLr isoform of 50 kDa (Figure 3A) and PRL of 60 kDa and 23 kDa were found (Figure 3B). Until 8 h after stimulation with LPS, the increased expression of PRLr isoforms of 50 kDa (p<0.05) and 100 kDa (p<0.001) (Figure 3A) and PRL of 60 kDa (p<0.01) was observed, as well as, the decrease of PRL of 23 kDa (p<0.01) (Figure 3B). After 48 hours of stimulation with LPS the increased expression of PRLr isoforms of 100, 90, 65 and 50 kDa (Figure 3A), and the increase of the 80 kD PRL (p<0.001) was revealed, as well as a decrease of PRL of 60 kDa (Figure 3B). PRL of 23 kDa was not detected after 8 h of stimulation with LPS (Figure 3B).