Journal: Frontiers in Molecular Neuroscience

Article Title: An IQSEC2 Mutation Associated With Intellectual Disability and Autism Results in Decreased Surface AMPA Receptors

doi: 10.3389/fnmol.2019.00043

Figure Lengend Snippet: Flow cytometry analysis of hippocampal derived cells for GluA1/2. (A) Representative dot plot of one of 10 experiments. US panel-unstained hippocampal cells. Mut and WT panels demonstrating staining with Alexa Flour 647 for extracellular GluA1/2 in single cell suspension of hippocampal cells from A350V and wild type mice, respectively, and window selected for identifying GluA1/2+ cells. (B) Summary of all experiments demonstrating significantly higher percentage of cells staining with GluA1/2 in wild type as compared to A350V mutant hippocampal neuronal preparations (4.1 ± 0.5 vs. 3.1 ± 0.5, paired t -test, n = 10 independent experiments, ∗ p < 0.00001).

Article Snippet: 20 ug of total protein from each sample was resolved on 10% SDS-PAGE and western immunoblotting was performed to analyze the surface and intracellular pools of AMPA receptors using anti-GluA1 (rabbit monoclonal, Abcam, Ab109450) (1:1,000), anti-GluA2 (rabbit monoclonal, Abcam Ab150387) (1:2,000), anti-GluA3 (rabbit monoclonal, Abcam Ab40845) (1:1,000), anti-GluA4 (goat polyclonal, Abcam Ab115322) (1:1,000), HRP conjugated polyclonal goat anti-rabbit (Cell signaling 7074), HRP conjugated polyclonal rabbit anti-goat (Invitrogen 611620) and HRP conjugated monoclonal mouse anti-beta-actin (Millipore-SIGMA, A3854) (1:5,000).

Techniques: Flow Cytometry, Derivative Assay, Staining, Mutagenesis

Journal: bioRxiv

Article Title: Riluzole does not ameliorate disease caused by cytoplasmic TDP-43 in a mouse model of amyotrophic lateral sclerosis

doi: 10.1101/749846

Figure Lengend Snippet: A Representative immunoblots of RIPA-soluble and urea-soluble cortical brain lysates from vehicle- or riluzole-treated control and rNLS mice for human-specific (h)TDP-43, human- and mouse-specific (h+m)TDP-43 and phosphorylated (p409/410) TDP-43. Quantification of B RIPA-soluble hTDP-43ΔNLS, C RIPA-soluble h+mTDP-43, D urea-soluble hTDP-43ΔNLS, E urea-soluble h+mTDP-43, and F p409/410 TDP-43 revealed higher levels in rNLS mice, but no effect of riluzole treatment. GAPDH for loading control of hTDP-43 and h+mTDP-43 blot is shown, with full blots and individual matched GAPDH/total protein blots used for quantification shown in . n =3/control group and n =5/rNLS group. Detailed statistical test results are described in .

Article Snippet: Primary antibodies included: rabbit anti-human/mouse TDP-43 1:5000 (Proteintech Cat# 10782-2-AP, RRID:AB_615042), mouse anti-human TDP-43 1:5000 (Clone 5104, ( )), mouse anti-phospho-TDP-43 (Ser409/410) (Cosmo Bio Co Cat# CAC-TIP-PTD-M01, RRID:AB_1961900), rabbit anti-EAAT2 1:1000 (Abcam Cat# ab178401), rabbit anti-GluA1 1:1000 (Abcam Cat# ab31232, RRID:AB_2113447), rabbit anti-GluA2 1:1000 (Millipore Cat# AB1768, RRID:AB_2247874), rabbit anti-GluA3 1:1000 (Abcam Cat# ab40845, RRID:AB_776310), and mouse anti-GAPDH 1:5000 (Proteintech Cat# 60004-1-Ig, RRID:AB_2107436), diluted in 0.1% BSA overnight at 4°C.

Techniques: Western Blot