ABSTRACT
The stability of reference proteins in semi-quantitative Western blot experiments in normal and diseased placenta has never been studied. This study aims to determine the stability of five reference proteins and two general protein stains in placentas from preeclampsia, gestational diabetes mellitus and matched control pregnancies. The stability of the reference proteins was analysed using indicators of inter-group (P value) and intra-group (coefficient of variation) stability. The effect of different normalization strategies was determined by normalizing serotonin transporter (SERT) expression against the different reference protein markers. Results show significant expression variability of ß-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1), peptidylprolyl isomerase A (PPIA) and α-tubulin, and that amido black staining is the most stable reference protein marker. Furthermore, results show that SERT expression significantly differs according to the reference protein markers used for its normalization. The present study demonstrated the importance of using stable reference protein markers and normalization strategy in order to get correct results in semi-quantitative Western blot experiments in placental tissues.
Subject(s)
Placenta/metabolism , Pregnancy Proteins/metabolism , Protein Stability , Staining and Labeling/standards , Adult , Case-Control Studies , Diagnostic Techniques, Obstetrical and Gynecological/standards , Female , Humans , Infant, Newborn , Placenta/pathology , Pregnancy , Pregnancy Complications/diagnosis , Pregnancy Complications/metabolism , Pregnancy Complications/pathology , Reference Standards , Young AdultABSTRACT
Human villous trophoblast differentiation is a complex and highly regulated process essential for the well-being of the pregnancy and fetal development. In this review, we present an overview of the role of MAPKs signalling in morphological and functional differentiation of villous trophoblast.
