Gestational Regulation of Granulocyte-Colony Stimulating Factor Receptor Expression: MATERIALS AND METHODS(3)

MATERIALS AND METHODS(3)

Total RNA was isolated from 11 first-trimester, 10 second-trimester, and 8 third-trimester placental tissues according to a standard procedure. Placental RNA (15 ^g) was subjected to RpA using the G-CSFR and GApDh probes and the HybSpeed RPA Kit (Ambion, Witney, UK) according to the manufacturer’s instructions. The protected RNA hybrids were subjected to 6% denaturing PAGE. Den-sitometric analysis was performed using BioImage Intelligent Quantifier software (ImproVison, Coventry, UK). Statistical analyses were performed using the Mann Whitney U-test for nonparametric variables, and p < 0.05 was considered statistically significant. buy cheap antibiotics

Western Blotting

Cell membrane preparations from 11 first-, 10 second-, and 8 third-trimester frozen placentas were processed from tissue homogenates and subjected to Western blotting using LM832 and anti-actin antibody (Sigma Chemical Company, St. Louis, MO) as previously described. Actin and G-CSFR were detected by the enhanced chemiluminescence (ECL) Western Blotting System (Amersham Life Science Ltd.) according to the manufacturer’s instructions. Densi-tometric analyses of G-CSFR and actin bands was performed as above.

This entry was posted in Human Placenta and tagged Human Placenta, Receptor Expression, Stimulating Factor.