Journal: bioRxiv

Article Title: Multiple Postsynaptic Protein Levels Are Maintained in a Delayed Form of Compensatory, Experience-Dependent Plasticity in Adult Superior Colliculus

doi: 10.1101/2022.10.06.511220

Figure Lengend Snippet: (A) Upper panel: Individual Western blots of normally reared (NR) and dark-reared (DR) treatment groups generated using 20 μg of SC protein per lane. Lower panel: Densitometric analyses of GABA A Rα2 in normally vs. dark reared hamster pups. (B) Comparison of GABA A Rα2 expression with GABAA A Rα1 expression, shown as individual Western blots in the upper panel and as a ratio in the lower panel. Data presented as mean ± SEM. In this and other figures, lanes presented together are from the same gel(s), and each measured protein was normalized against GAPDH as a loading control. These results suggest that the number and ratio of synaptic GABA A α2 receptors are similar in normal and dark reared adult SC.

Article Snippet: Primary antibodies used in this study included: rabbit anti-GABA A Rα1 (1:1000, Cat#: AGA-001, Alomone Labs); rabbit anti-GABA A Rα2 (1:1000, Cat#: ab72445, Abcam); rabbit anti-GABA A Rα5 (1:1000, Cat#: ab10098, Abcam); rabbit anti-Gephyrin (1:1000, Cat#: ab32206, Abcam); mouse anti-PSD-95 (1:1000, Cat#: ab2723, Abcam); mouse anti-KCC2 (1:1000, Cat#: 75-013, NeuroMab); rabbit anti-NKCC1 (1:1000, Cat#: ab59791, Abcam); rabbit anti-NMDAR2A (1:1000, Cat#: ab133265, Abcam); mouse anti-NMDAR2B (1:1000, Cat#: ab93610, Abcam); and mouse anti-GAPDH (1:1000, Cat#: 600-GAPDH, PhosphoSolutions).

Techniques: Western Blot, Generated, Comparison, Expressing, Control

Journal: bioRxiv

Article Title: Multiple Postsynaptic Protein Levels Are Maintained in a Delayed Form of Compensatory, Experience-Dependent Plasticity in Adult Superior Colliculus

doi: 10.1101/2022.10.06.511220

Figure Lengend Snippet: Example blots of NR and DR treatment groups as in . All presented lanes are from the same gel(s), and each measured protein was normalized against GAPDH as a loading control. (A) GABA A Rα5 comparison. (B) GABA A Rα1: GABA A Rα5 ratio. All presented lanes are from the same gel(s), and each measured protein was normalized against GAPDH as a loading control. Data presented as mean ± SEM. These results suggest that the number and ratio of synaptic GABA A α5 receptors are similar in normal and dark reared adult SC.

Article Snippet: Primary antibodies used in this study included: rabbit anti-GABA A Rα1 (1:1000, Cat#: AGA-001, Alomone Labs); rabbit anti-GABA A Rα2 (1:1000, Cat#: ab72445, Abcam); rabbit anti-GABA A Rα5 (1:1000, Cat#: ab10098, Abcam); rabbit anti-Gephyrin (1:1000, Cat#: ab32206, Abcam); mouse anti-PSD-95 (1:1000, Cat#: ab2723, Abcam); mouse anti-KCC2 (1:1000, Cat#: 75-013, NeuroMab); rabbit anti-NKCC1 (1:1000, Cat#: ab59791, Abcam); rabbit anti-NMDAR2A (1:1000, Cat#: ab133265, Abcam); mouse anti-NMDAR2B (1:1000, Cat#: ab93610, Abcam); and mouse anti-GAPDH (1:1000, Cat#: 600-GAPDH, PhosphoSolutions).

Techniques: Control, Comparison

Journal: bioRxiv

Article Title: Multiple Postsynaptic Protein Levels Are Maintained in a Delayed Form of Compensatory, Experience-Dependent Plasticity in Adult Superior Colliculus

doi: 10.1101/2022.10.06.511220

Figure Lengend Snippet: ( A ) Adult levels of the cytosolic vs. membrane-attached ratio of the extrasynaptically-targeted GABA A Rα5 and (B) the synaptically-targeted GABA A Rα1 subunits were not affected by early light deprivation. Example blots (upper panels) represent bands of labeled intracellular and membrane bound receptor subunit proteins (upper bands) from the same animal, measured as a ratio and compared between NR and DR groups. GAPDH was used as a loading control. Graph data presented as mean relative optical density ± SEM.

Article Snippet: Primary antibodies used in this study included: rabbit anti-GABA A Rα1 (1:1000, Cat#: AGA-001, Alomone Labs); rabbit anti-GABA A Rα2 (1:1000, Cat#: ab72445, Abcam); rabbit anti-GABA A Rα5 (1:1000, Cat#: ab10098, Abcam); rabbit anti-Gephyrin (1:1000, Cat#: ab32206, Abcam); mouse anti-PSD-95 (1:1000, Cat#: ab2723, Abcam); mouse anti-KCC2 (1:1000, Cat#: 75-013, NeuroMab); rabbit anti-NKCC1 (1:1000, Cat#: ab59791, Abcam); rabbit anti-NMDAR2A (1:1000, Cat#: ab133265, Abcam); mouse anti-NMDAR2B (1:1000, Cat#: ab93610, Abcam); and mouse anti-GAPDH (1:1000, Cat#: 600-GAPDH, PhosphoSolutions).

Techniques: Membrane, Labeling, Control

Journal: PLoS ONE

Article Title: Interleukin-22 Suppresses the Growth of A498 Renal Cell Carcinoma Cells via Regulation of STAT1 Pathway

doi: 10.1371/journal.pone.0020382

Figure Lengend Snippet: Western blot assay was performed to detect the expression of IL-22R on the surface of cells using mouse IL-22 Rα1 antibody. The results showed a measurable expression of IL-22R on the surface of A498 cells compared with that of HepG2 cells and human B cells, which served as positive control and negative control respectively. N = 3.

Article Snippet: Mouse IL-22 Rα1 antibody was purchased from R&D system.

Techniques: Western Blot, Expressing, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: Kinetic Analysis of Mouse Brain Proteome Alterations Following Chikungunya Virus Infection before and after Appearance of Clinical Symptoms

doi: 10.1371/journal.pone.0091397

Figure Lengend Snippet: Top-5 networks from IPA of the total 177 proteins differentially expressed identified by 2D-DIGE and iTRAQ labeling following CHIKV infection, considering the 3 comparisons of early (E) vs mock (M), late paralytic (LP) or late tetanus-like (LT) vs E samples.

Article Snippet: Blots were saturated with 5% nonfat dried milk in PBS containing 0.05% (v/v) Tween 20 (PBS-T-milk) for 1 h. Western blot (WB) analyses were carried out with rabbit mono- or polyclonal antibodies directed against β-arrestin (1∶5000, ARRB1, no. 4674, Cell Signaling Technology, Danvers, MA), GRIP-associated protein (1∶500, GRASP1, no. sc-135681, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), annexin A2 (1∶100, ANXA2, no. sc-9061, Santa Cruz), integrin αV (1∶100, ITGAV, no. 10179, Santa Cruz), myosin phosphatase target subunit 1 (1∶100, MYPT1, no. sc-25618, Santa Cruz), rabaptin-5 (1∶1000, RABEP1, no. sc-15351, Santa Cruz), N-Ras (1∶500, N-Ras, no. sc-519, Santa Cruz), synaptogyrin-3 (1∶1000, SYNGR3, no. sc-68936, Santa Cruz), or with a goat polyclonal antibody directed against γ-aminobutyric acid receptor subunit alpha-1 (1∶100, GABA A Rα1 or GABRA1, no. sc-31045, Santa Cruz), diluted in PBS-T-milk and incubated overnight at 4°C.

Techniques: Multiplex sample analysis, Labeling, Infection, Ubiquitin Proteomics, Modification

Journal: PLoS ONE

Article Title: Kinetic Analysis of Mouse Brain Proteome Alterations Following Chikungunya Virus Infection before and after Appearance of Clinical Symptoms

doi: 10.1371/journal.pone.0091397

Figure Lengend Snippet: (A) Protein samples from each group used for proteomic analysis were minimally labeled with cyanine-3 dye. At the top, a representative protein profile of three biological replicates from brain lysates of mock (M), early (E), late paralytic (LP) and late tetanus-like (LT), separated by 10% SDS-PAGE is shown. WB with fluorescence-based methods was used to detect an overlaid fluorescent scan of the general protein patterns (Cy3 dye; green) and the specific immunoreactive proteins (FITC or Cy5 dye; red). To better visualize protein detection signals observed with each specific antibody used, corresponding cropped WB images are presented in grey levels. (B) The graphs correspond to the mean ± S.D. of protein quantity measured by densitometry of the antigenic bands. Densitometry analyses were performed using TotalLab Quant v12.2 software (Nonlinear Dynamics), and data were normalized to levels of global protein pattern intensity. The values indicated under each graph correspond to fold changes from paired comparisons. The significance of the differential protein expression are indicated *, p<0.05; **, p<0.01; ***, p<0.001. A.U., arbitrary units. ANXA2: annexin A2; ARRB1: β-arrestin; GABRA1: γ-aminobutyric acid receptor subunit alpha-1; GRASP1: GRIP-associated protein; ITGAV: integrin αV; MYPT1: myosin phosphatase target subunit 1; N-Ras: N-Ras; RABEP1: rabaptin-5; SYNGR3: synaptogyrin-3.

Article Snippet: Blots were saturated with 5% nonfat dried milk in PBS containing 0.05% (v/v) Tween 20 (PBS-T-milk) for 1 h. Western blot (WB) analyses were carried out with rabbit mono- or polyclonal antibodies directed against β-arrestin (1∶5000, ARRB1, no. 4674, Cell Signaling Technology, Danvers, MA), GRIP-associated protein (1∶500, GRASP1, no. sc-135681, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), annexin A2 (1∶100, ANXA2, no. sc-9061, Santa Cruz), integrin αV (1∶100, ITGAV, no. 10179, Santa Cruz), myosin phosphatase target subunit 1 (1∶100, MYPT1, no. sc-25618, Santa Cruz), rabaptin-5 (1∶1000, RABEP1, no. sc-15351, Santa Cruz), N-Ras (1∶500, N-Ras, no. sc-519, Santa Cruz), synaptogyrin-3 (1∶1000, SYNGR3, no. sc-68936, Santa Cruz), or with a goat polyclonal antibody directed against γ-aminobutyric acid receptor subunit alpha-1 (1∶100, GABA A Rα1 or GABRA1, no. sc-31045, Santa Cruz), diluted in PBS-T-milk and incubated overnight at 4°C.

Techniques: Labeling, SDS Page, Fluorescence, Software, Expressing

Journal: Frontiers in Pharmacology

Article Title: The Effects of Ginsenoside Compound K Against Epilepsy by Enhancing the γ-Aminobutyric Acid Signaling Pathway

doi: 10.3389/fphar.2018.01020

Figure Lengend Snippet: Effects of GCK on the expression of GABA A Rα1 and NMDAR1 protein in the hippocampus of the pilocarpine-induced epileptic rats. (A) The expression of GABA A Rα1 in the Con, Pilo, VPA, and GCK-treated groups. (B) The expression of NMDAR1 in the Con, Pilo, VPA, and GCK-treated groups. (C–G) The GABA A Rα1 immunoreactivity in the hippocampal CA1, CA3, DG, and H regions. Scale bar: 100 μm. Values are mean ± SEM ( n = 5). Pilo 30 mg/kg, VPA 400 mg/kg, GCK 80 mg/kg, 160 mg/kg, and 320 mg/kg. Compared with the Con ∗ P < 0.05; Compared with the Pilo, # P < 0.05.

Article Snippet: Sections were incubated overnight at 4°C with the primary antibodies: the mouse anti-GABA A Rα1 monoclonal antibody (1:100, Abcam, United Kingdom), the goat anti-NKCC1 polyclonal antibody (1:50, Santa Cruz, United States), and the goat anti-KCC2 polyclonal antibody (1:50, Santa Cruz, United States).

Techniques: Expressing

Journal: Cell reports

Article Title: Local miRNA-Dependent Translational Control of GABA A R Synthesis during Inhibitory Long-Term Potentiation

doi: 10.1016/j.celrep.2020.107785

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-puromycin (Kerafast 3RH11) and anti-GABA A Rα1 (Synaptic systems 224203) were used at 1:500.

Techniques: Imaging, Recombinant, Reporter Assay, Software, Western Blot