Journal: The Journal of Neuroscience

Article Title: β1-C121W Is Down But Not Out: Epilepsy-AssociatedScn1b-C121WResults in a Deleterious Gain-of-Function

doi: 10.1523/jneurosci.0405-16.2016

Figure Lengend Snippet: Figure2. Comparisonof1-C121WandWT1proteinexpression.A,RepresentativeWesternblotscomparinganti-1 immunoreactivity in Scn1b / and Scn1b W/W mouse brain membrane preparations. 1-C121W polypeptides migrate at a lower apparent molecular weight than WT 1 separated on a 10% SDS-PAGE gel. Immunoreactive bands were quantified using densitometry. Each band density was first normalized to its corresponding -tubulin signal and 1 levels in Scn1b W/Wmice were expressed as a percentage of 1 in Scn1b /mice. Scn1b W/Wmice had 45 10% of Scn1b /1 expression (2-way ANOVA; p 0.0001; 3 replicate experiments with 4 animals of each genotype). B, Representative Western blots showing mock-digested, PNGaseF-digested, and control brain membrane inputs from Scn1b / and Scn1b W/W brains separated on a 12% SDS-PAGE gel. WT 1 and 1-C121W polypeptides migrate at similar molecular weights after removal of N-linked glycosylation by PNGaseF (n 3). C, Representative Western blots, separated on a 15% SDS-PAGE gel, comparing levels of WT 1 and 1-C121W polypeptides in P14–P15, P17, and 20-week-old Scn1b /, Scn1b /W, and Scn1b W/W mice. -tubulin is shown as a loading control. Every Scn1b W/W mouse tested showed the lower molecular band only (P14–P15, n 5; P17, n 5). Every Scn1b /W sample tested showed both bands (P14–P15, n 11; P17, n 6; 20 weeks, n 4).

Article Snippet: Primary antibodies were as follows: rabbit anti-Scn1b (directed against an intracellular 1 epitope; Cell Signaling Technology, preproduction serum of D4Z2N, catalog #14684, 1:3000 used for Western blotting); rabbit anti-Scn1b (directed against an extracellular 1 epitope; Cell Signaling Technology, preproduction serum of D9T5B, catalog #13950, 1:25 used for immunofluorescence in optic nerves); rabbit anti-Scn1b (directed against an extracellular 1 epitope; Cell Signaling Technology, production version of D9T5B, catalog #13950, 1:250 used for immunofluorescence in brains); guinea pig anti-Caspr (gift from Dr. James Salzer, New York University School of Medicine, 1:1000 used for immunofluorescence in optic nerves); mouse anti-PAN VGSC -subunit (Sigma-Aldrich, catalog #S8809, 1:200 used for immunofluorescence in optic nerves and coimmunoprecipitation); goat anti-ankyrinG (recognizing total ankyrinG, gift from Dr. Vann Bennett, He et al., 2014; Jenkins et al., 2015, 1:500 used for immunofluorescence in brains); mouse anti-calbindin (Sigma-Aldrich, catalog #C9848, 1:400 used for immunofluorescence in brains); rat anti-Ctip2 (Abcam, catalog #ab18465, 1:400 used for immunofluorescence in brains); rabbit anti-PAN VGSC -subunit (Cell Signaling Technology, D2I9C, catalog #14380, 1:1000 used for Western blotting); mouse anti-human -tubulin (Cedarlane Laboratories, catalog #CLT9002; 1:5000, used for Western blotting); mouse anti-HSP90 (Enzo Life Sciences, catalog #AC88, 1:1000, used for Western blotting).

Techniques: Membrane, Molecular Weight, SDS Page, Expressing, Western Blot, Control, Glycoproteomics

Journal: Endocrinology

Article Title: In utero exposure to the endocrine disruptor di-(2-ethylhexyl) phthalate induces long-term changes in gene expression in the adult male adrenal gland.

doi: 10.1210/en.2013-1921

Figure Lengend Snippet: Figure 6. Serum markers that correlate with in utero exposure to DEHP. A, Heatmap of differentially expressed genes at PND21 and PND60 in offspring of rats exposed in utero to 100- or 300-mg DEHP/kgd was constructed to identify genes altered at both time points and doses. Selected gene targets (bold) were used to evaluate putative serum markers. Serum levels of AQP7 (B), FABP4 (C), and PCK1 (D) in adult rats exposed in utero to the indicated doses of DEHP. Data in C and D are means SD (n 4); **, ANOVA, P .01. E, Relationship between FABP4 and PCK1 adult rat serum levels to in utero DEHP exposure doses used. FABP4, correlation coefficient 0.37, P .49. PCK1, correlation coefficient 0.77, P .1.

Article Snippet: ELISAs were used to quantify aquaporin 7 (AQP7) (catalog number CSB-EL001967RA; Cusabio Biotech, Cedarlane Laboratories), fatty acid-binding protein 4 (FABP4) (catalog number CSB-E13027r; Cusabio Biotech), and phosphoenolpyruvate carboxykinase I (PCK1) (catalog number sE90936Ra; USCN Life Science, Inc, Cedarlane Laboratories), according to the instructions of each manufacturer.

Techniques: In Utero, Construct