Journal: Molecular human reproduction

Article Title: Decreased expression of NR4A nuclear receptors in adenomyosis impairs endometrial decidualization.

doi: 10.1093/molehr/gaw042

Figure Lengend Snippet: Figure 2 NR4A regulation of human endometrial stromal cell (hESC) decidualization in vitro. hESCs were transduced with Ad-LacZ or Ad-NR4A at a multiplicity of infection (MOI) of 25 or were treated with 0.5 mM 8-Br-cAMP and 1 µM MPA for 48 h. PRL (A) and IGFBP1 (B) mRNA levels were measured by real-time PCR. The results are presented as the mean ± SEM; n = 3 samples. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with Ad-LacZ. hESCs were infected with the indicated adenoviruses at a MOI of 50, followed by treatment with or without 8-Br-cAMP and MPA for an add- itional 4 days. Prolactin (C) and IGFBP-1 (D) released into the medium was measured by Enzyme Linked Fluorescent Assay (ELFA) and ELISA. The results are presented as the mean ± SEM; n = 5 samples. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with Ad-LacZ. hESCs were transfected with 50 nM of the indicated specific NR4A siRNA for 72 h, followed by treatment with or without 8-Br-cAMP and MPA for an additional 4 days. Prolactin (E) and IGFBP-1 (F) released into the medium were measured by ELFA and ELISA. The results are presented as the mean ± SEM; n = 4 sam- ples. *** P < 0.001 compared with siCTL. # P < 0.05 and ## P < 0.01 compared with siCTL plus 8-Br-cAMP and MPA. (G and H) Fluorescein isothio- cyanate labeled phalloidin was used to label actin filaments, and immunofluorescence was used to analyze the morphological transformation of hESCs.

Article Snippet: Detection of prolactin and IGFBP-1 levels in cultured supernatants Prolactin and IGFBP-1 levels in supernatants were measured using a Vidas prolactin kit (bioMérieux) and a human IGFBP1 ELISA kit (BOSTER).

Techniques: In Vitro, Transduction, Infection, Real-time Polymerase Chain Reaction, Fluorescence, Enzyme-linked Immunosorbent Assay, Transfection, Labeling, Transformation Assay

Journal: Lab on a chip

Article Title: Point-of-Care Diagnostics to Improve Maternal and Neonatal Health in Low-Resource Settings

doi: 10.1039/c7lc00374a

Figure Lengend Snippet: Representative commercially available technologies available to diagnose the leading causes of maternal mortality. Notably, many of the listed technologies are lacking robust usability, scalability, and field performance studies.

Article Snippet: The two most commonly reviewed tests are the Amnisure ROM test, which detects the biomarker placental alpha-microglobulin 1 (PAMG-1), and the Actim PROM tests which identifies the presence of Insulinlike growth factor binding protein 1 (IGFBP-1).

Techniques: Diagnostic Assay, Biomarker Discovery, Infection, High Throughput Screening Assay, Sample Prep, Clinical Proteomics, Sterility

Journal: Genomics, Proteomics & Bioinformatics

Article Title: Oleic Acid and Eicosapentaenoic Acid Reverse Palmitic Acid-induced Insulin Resistance in Human HepG2 Cells via the Reactive Oxygen Species/ JUN Pathway

doi: 10.1016/j.gpb.2019.06.005

Figure Lengend Snippet: SILAC-based quantitative proteomic analysis A. Flow diagram of SILAC labeling combined with LC–MS/MS. B. and C. Quantitation overlap of the triplicate SILAC-based quantitative proteomic experiments upon PA+OA treatment (B) or PA+EPA treatment (C). Forward 1 and Forward 2 represent forward SILAC labeling experiments; Reverse 1 represents reverse SILAC labeling experiment. D. Western blots showing the expression of Jun and IGFBP1 in HepG2 cells treated with PA+OA treatment. E. SILAC quantitative proteomic ratios of Jun and IGFBP1 in HepG2 cells treated with PA+OA. F. Western blots showing the expression of six proteins in HepG2 cells treated with PA+EPA. G. SILAC quantitative proteomic ratios of six proteins in HepG2 cells treated with PA+EPA. SILAC, stable isotope labeling with amino acids in cell culture; RPLC, reversed-phase liquid chromatography; LC–MS/MS, liquid chromatography–tandem mass spectrometry; IGFBP1, insulin-like growth factor-binding protein 1; ATP5B, mitochondrial ATP synthase subunit β; EPCAM, epithelial cell adhesion molecule; HINT2, histidine triad nucleotide-binding protein 2; CYCS, cytochrome c; CDK1, cyclin-dependent kinase 1.

Article Snippet: To validate the OA-treated SILAC results, HepG2 cells were treated with ethanol, 0.5 mM PA, and 0.5 mM PA plus 0.2 mM OA for 12 h, respectively, and then treated with 100 nM insulin for 20 min. Proteins were then extracted, and Western blots were performed to detect the expression levels of Jun (1:1000; Catalog No. 9165T, Cell Signaling Technology) and IGFBP1 (1:1000; Catalog No. 31025T, Cell Signaling Technology).

Techniques: Multiplex sample analysis, Labeling, Liquid Chromatography with Mass Spectroscopy, Quantitation Assay, Western Blot, Expressing, Quantitative Proteomics, Cell Culture, Reversed-phase Chromatography, Liquid Chromatography, Mass Spectrometry, Binding Assay