Journal: Placenta

Article Title: Placental Glucose transporter 3 (GLUT3) is Up-regulated in Human Pregnancies Complicated by Late-onset Intrauterine Growth Restriction

doi: 10.1016/j.placenta.2013.08.010

Figure Lengend Snippet: Immunohistochemical localization of GLUT3 and HIF-1α in human placenta. (A) Controls for GLUT3 staining at 600x magnification. GLUT3 staining of human testis; there was no GLUT3 staining in sections where the primary antibody was omitted (A1). Positive control (testis) shows immunoreaction for GLUT3 (brown) in the maturing secondary spermatocytes and sperm (A2). (B) Sections of three different subjects showing normal term placenta (B1-B3) vs. IUGR (slide B4=sample 8, slide B5=sample 10, slide B6=sample 5), stained with GLUT3, staining at 400x magnification. GLUT3 is localized to the cytotrophoblast layer and the syncytiotrophoblast layer to a lesser degree. Arrows indicate immunoreactive syncytium, as determined by their morphology and location relative to the intervillous space. Triangles show underlying cytotrophoblast layer. Scale bars = 50μm (C) GLUT3 positive staining area per squared micrometers was quantified using Dynamiccellcount BZ-HIC software. Digital analysis revealed increased GLUT3 immunopositivity in IUGR as compared to normal placenta. *P < 0.05 (D) Double immunohistochemical localization of GLUT3(red) and HIF -1α(green) in control placenta (slides D1-D4) and IUGR (slides D5-D8). D1 and D5: stained for GLUT3 (Texas Red) and DAPI (nuclear blue). D2 and D6: stained for HIF-1α (FITC green) and DAPI (nuclear blue). D3 and D7: triple stained for GLUT3 (Texas Red), HIF-1α (FITC green), and DAPI (nuclear blue). Arrows indicate immunoreactive syncytium, as determined by their morphology and location relative to the intervillous space. D4 and D8: negative controls performed by omission of primary antibody and showed that cross-staining did not occur. Scale bars = 50μm (E) HIF-1α (green) staining in normal placenta (E1) vs. IUGR, fetal aspect (E2) vs. IUGR, maternal aspect (E3). Arrows indicate immunoreactive syncytium. Scale bars = 50μm.

Article Snippet: Blots were probed in the following primary antibodies: rabbit-polyclonal for GLUT1 (Santa Cruz), GLUT4 (ABcam) HIF-1α (Cell Signaling), and mouse monoclonal for GLUT3 (Santa Cruz).

Techniques: Immunohistochemical staining, Staining, Positive Control, Software

Journal: Placenta

Article Title: Placental Glucose transporter 3 (GLUT3) is Up-regulated in Human Pregnancies Complicated by Late-onset Intrauterine Growth Restriction

doi: 10.1016/j.placenta.2013.08.010

Figure Lengend Snippet: (A) Real-time quantitative RT-PCR analysis of placental GLUT1, GLUT3 and GLUT4 expression. IUGR placentas (n=10) are assayed against gestationally age-matched, control placentas (n=10) for the maternal vs. fetal side. Relative quantification of PCR products was based on the Ct value differences between target and the housekeeping gene using the comparative Ct method (Eukaryotic 18s rRNA (Applied Biosystems) was used as an internal control. *P < 0.05. (B) Western blot analysis of GLUT1, GLUT3, and GLUT4 in placental biopsies obtained from women with normal pregnancy (n=10) or pregnancy affected by IUGR, suspected by abnormal prenatal ultrasound (n=10). Protein taken from fetal (F) and maternal (M) sides were loaded on 12% SDS-polyacrylamide gels and transferred onto PVDF membranes. Representative blots are shown. (B, Bottom) Arbitrary densitometric units showing mean of all samples; data are normalized to a control value of 100%, bars indicate SE. *P < 0.05.

Article Snippet: Blots were probed in the following primary antibodies: rabbit-polyclonal for GLUT1 (Santa Cruz), GLUT4 (ABcam) HIF-1α (Cell Signaling), and mouse monoclonal for GLUT3 (Santa Cruz).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Placenta

Article Title: Placental Glucose transporter 3 (GLUT3) is Up-regulated in Human Pregnancies Complicated by Late-onset Intrauterine Growth Restriction

doi: 10.1016/j.placenta.2013.08.010

Figure Lengend Snippet: DNA methylation analysis of placental GLUT3 promoter regions in placental biopsies obtained from the maternal and fetal sides in women with normal pregnancy and pregnancy affected by IUGR. Genomic DNA was processed by pyrosequencing.

Article Snippet: Blots were probed in the following primary antibodies: rabbit-polyclonal for GLUT1 (Santa Cruz), GLUT4 (ABcam) HIF-1α (Cell Signaling), and mouse monoclonal for GLUT3 (Santa Cruz).

Techniques: DNA Methylation Assay