The data obtained from three independent experiments were utilized for analysis of relative gene expression. SD. The relative intensity of CD81 levels was assessed by Image J. *< 0.05, **< 0.01. Open in a separate windows Fig. S1. Vimentin, CD45, and CD66 immunostainings are expressed in the 19-wk villous core of FV. (= 5. (Level bar, 25 m.) To determine whether CD81 expression is usually regulated at the transcriptional or translational level in CTBs isolated from first, second, and third TM placentas, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting analyses were used to quantify the levels of CD81 mRNA and protein. Compared with the CTBs from your first TM villi, lower CD81 mRNA and protein expression levels were observed in CTBs from the second TM placentas, and CD81 protein expression was significantly down-regulated in CTBs from the third TM placentas (Fig. 1 and and Fig. S2and Fig. S2and = 15) and nPTB (= 15) (sPE vs. nPTB, < 0.01). (and = 3 in each group; sPE vs. nPTB, < 0.05). (and = 12) and gestational age-matched nPTB (= 12) (sPE vs. nPTB, < 0.01). (and < 0.01; = 12 in each group). (and < 0.05; = 12 in each group). All Western EG01377 TFA blotting data are offered as mean SD. The relative intensity of CD81 levels was assessed by Image J. GA, gestational age. *< 0.05, **< 0.01. Open in a separate windows Fig. S2. CD81 expression is usually up-regulated on CTBs, STBs, and the cells of the villous core from patients EG01377 TFA with early-onset sPE. (= 5). (= 5). (Level bar, 25 m.) Because CD81 expression was significantly up-regulated in the sPE placentas, we examined whether these placentas released increased levels of CD81 protein into maternal blood circulation. A total of 24 serum samples were collected, including 12 from patients with early-onset sPE and 12 from gestational age-matched nPTB. An immunoblotting analysis showed that serum CD81 levels were significantly increased in the patients with sPE (Fig. 2 and and and and and < 0.01). (< 0.05 Rabbit Polyclonal to TSN and < EG01377 TFA 0.01). (Level bar, 500 m.) (and < 0.05, **< 0.01. The principal pathogenesis of PE is usually thought to be associated with maternal endothelial cell activation (27). In an attempt to understand the pathogenic role of CD81 in sPE, we tested the hypothesis that increased CD81 levels in maternal blood circulation cause endothelial lesions. We performed a tube formation assay with HUVECs using recombinant CD81. As shown in Fig. 3 and < 0.05, **< 0.01.) CD81-Induced Rat Model Mimics the Phenotype of Human PE. In previous studies, a low-dose infusion EG01377 TFA of LPS into rats at the early stages of pregnancy successfully triggers a PE-like phenotype (28C30). Inspired by this model, we tested the hypothesis that CD81 overproduction participates in preeclamptic placentation and triggers the clinical manifestations of PE. Pregnant rats were infected with either Ad-CD81 or Ad-CTL around the 5th day of gestation (GD5). As controls, nonpregnant rats were also infected on the same day. The blood pressure and proteinuria of the rats were monitored, and both groups of rats were euthanized either 10 or 14 d after contamination (at GD15 or GD19). CD81 expression and trophoblast-directed uterine spiral artery remodeling were also analyzed. Compared with the Ad-CTLCinfected rats, the Ad-CD81Cinfected rats exhibited a significant elevation in systolic blood pressure (SBP; 113.5 1.95 mmHg vs. 108.76 4.62 mmHg) on GD6, and this elevation was maintained until GD14. Interestingly, Ad-CD81Cinfected nonpregnant rats showed no obvious changes in SBP during the same time period (Fig. 4< 0.01). Compared with the Ad-CTLCinfected rats,.