rabbit polyclonal usp10 antibody  (Bethyl)

Journal: iScience

Article Title: Chronic stress antagonizes formation of stress granules

doi: 10.1016/j.isci.2025.114556

Figure Lengend Snippet: Chronic stress-pre-incubated cells fail to disassemble polysomes by acute stress U2OS cells were subjected to treatment with 100 μM SA for 1 h after pre-incubation with 10 μM SA, 1 μM Tg (thapsigargin), 60 μM CCCP (carbonyl cyanide m -chlorophenyl hydrazone), or HBSS for 24 h. Unstressed cells (NT) were used as a control. (A) Cells were pulsed with puromycin and emetine for 5 min and lysed. Cell lysates were subjected to western blotting using antibodies for puromycin, p -eIF2α, total eIF2α, G3BP1, G3BP2, caprin1, USP10, and β-actin. Three blots were taken for each of the three biological replicates (independent experiments). The same colors of the circle dot in the graph are from the same independent experiment. Results are mean ± S.E.M. ( n = 3). p values were assessed using a one-way ANOVA ( p ∗ <0.05, p ∗∗ <0.01, p ∗∗∗ <0.001, p ∗∗∗∗ <0.0001). (B–F) Polysome profiles from U2OS cells. NT, black; 1 h SA, red; (B) 24 h pre-incubation of SA, blue; 1 h SA with 24 h pre-incubation of SA, green. (C) 24 h pre-incubation of Tg, blue; 1 h SA with 24 h pre-incubation of Tg, green. (D) 24 h pre-incubation of CCCP, blue; 1 h SA with 24 h pre-incubation of CCCP; green. (E) 24 h pre-incubation of HBSS, blue; 1 h SA with 24 h pre-incubation of HBSS; green. (F) The polysome/monosome ratio was calculated with normalization by each of NT as a control.

Article Snippet: Rabbit Polyclonal USP10 antibody , Bethyl Laboratories Inc. , A300-900A; RRID: AB_625312.

Techniques: Incubation, Control, Western Blot

Journal: eLife

Article Title: Processing of the ribosomal ubiquitin-like fusion protein FUBI-eS30/FAU is required for 40S maturation and depends on USP36

doi: 10.7554/eLife.70560

Figure Lengend Snippet: .

Article Snippet: Antibody , anti-USP10 (rabbit polyclonal) , Sigma-Aldrich , Cat# HPA006731 RRID: AB_1080495 , WB (1:1000).

Techniques: Cell Culture, Recombinant, Molecular Cloning, Sequencing, Northern Blot, Fluorescence, In Situ Hybridization, Negative Control

Journal: Cell

Article Title: G3BP1 is a tunable switch that triggers phase separation to assemble stress granules

doi: 10.1016/j.cell.2020.03.046

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-USP10 antibody , Proteintech , 19374–1-AP; RRID: AB_10858617.

Techniques: Virus, Recombinant, Protease Inhibitor, Electron Microscopy, Lysis, Extraction, Staining, Isolation, Mutagenesis, Sequencing, Plasmid Preparation, Software, Imaging

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: Tau is recruited in SGs in neuronal cells. ( a – d ) HT22 cells were treated with 5 µM MG-132 for 4 h and stained with anti-Tau (TAU-5), anti-TIA1, anti-USP10, or anti-G3BP. Nuclei were stained with Hoechst in ( c ). Staining was visualized by a fluorescent microscope. Arrowheads indicate the colocalization of Tau with both TIA1 and USP10 and Tau with G3BP. SG-positive cells were counted as described in the Materials and Methods section, and the population (%) of SG-positive cells is presented as the mean and SD from three independent experiments in ( b , d ). ** P < 0.01; *** P < 0.001. Scale bar: 10 µm. ( e , f ) Rat primary neuron-enriched cells were treated with 2 µM MG-132 for 8 h, and cells were stained with anti-Tau (TAU-5), anti-TIA1 and anti-USP10. SG-positive cells were counted, and the population (%) of SG-positive cells is presented as the mean and SD from three independent experiments in ( f ). * P < 0.05.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Staining, Microscopy

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: USP10-knockdown reduces Tau/TIA1-SG in HT22 cells. ( a ) Knockdown of USP10 (USP10-KD) in HT22 cells was performed using the lentivirus vector encoding USP10/shRNA ( USP10-3 , USP10-4 ) or the control ( NT ). The expression of USP10, Tau, TIA1 and β-actin in these cells was measured by Western blotting using respective antibodies. The asterisk indicates a nonspecific band. ( b , c ) USP10-KD HT22 cells and the control ( NT ) were treated with 5 µM MG-132 for 4 h, and SG formation was evaluated by anti-Tau (TAU-5) and anti-TIA1 antibodies as well as Hoechst for nuclear staining using a fluorescent microscope. Arrowheads indicate the colocalization of Tau with TIA1. The population (%) of cells with TIA1-SGs or TIA1/Tau-SGs is presented as the mean and SD from three independent experiments in ( c ). * P < 0.05; ** P < 0.01. Scale bar: 10 µm.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Plasmid Preparation, shRNA, Expressing, Western Blot, Staining, Microscopy

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: USP10-KD reduced the TIA1-induced Tau/TIA1-SG formation. ( a – c ) USP10-KD HT22 cells and the control ( NT ) were transfected with GFP-TIA1 plasmid using lipofection with Lipofectamine 2000. At 48 h after transfection, cells were treated with 5 µM MG-132 for 4 h, and SG formation was evaluated by GFP fluorescence together with anti-Tau (TAU-5) and anti-USP10 staining using a fluorescent microscope. Arrowheads indicate the colocalization of TIA1, Tau and USP10. The population (%) of cells with TIA1/Tau-SG or TIA1/Tau/USP10-SGs is presented as the mean and SD. ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar: 10 µm.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Transfection, Plasmid Preparation, Fluorescence, Staining, Microscopy

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: The deubiquitinase activity of USP10 is dispensable for Tau-SG formation. ( a ) A schematic illustration of USP10 and its mutants. ( b – d ) HT22 cells were transfected with a plasmid encoding HA-USP10 or its mutants (HA-USP10 C424A , HA-USP10 1–274 , HA- USP10 275–798 ) by lipofection using Lipofectamine 2000. At 48 h after transfection, SG formation was evaluated by anti-Tau (TAU-5), anti-TIA1 and anti-HA antibodies using a fluorescent microscope. Arrowheads indicate the colocalization of Tau, TIA1 and USP10. The population (%) of cells with Tau/TIA1/USP10-SGs is presented as the mean and SD from three experiments in ( d ). ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar: 10 µm. ( c ) The expression of USP10 and its mutants, Tau and β-actin in these cells was measured by Western blotting using anti-HA, anti-Tau (TAU-5) and anti-β-actin antibodies, respectively.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Activity Assay, Transfection, Plasmid Preparation, Microscopy, Expressing, Western Blot

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: MG-132 treatment disrupts the microtubule structure in HT22 cells. ( a – d ) HT22 cells were transfected with the HA-USP10 plasmid or control plasmid (HA) by the lipofection method using Lipofectamine 2000. At 48 h after transfection, cells were treated with 5 µM MG-132 or DMSO for 4 h. The localization of Tau and α-tubulin was evaluated by anti-Tau (TAU-5) and anti-α-tubulin antibodies as well as Hoechst using a fluorescence microscope. Arrows and arrowheads indicate Tau/α-tubulin aggregates and Tau aggregates, respectively. Cells with either Tau aggregates, α-tubulin aggregates or Tau/α-tubulin aggregates were counted, and the respective population (%) is presented as the mean and SD from three independent experiments in ( b – d ). ** P < 0.01. NS indicates that a result is not statistically significant. Scale bar: 10 µm.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Transfection, Plasmid Preparation, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: USP10 does not interact with Tau. ( a ) HT22 cells were transfected with Tau (Tau4R1N) plasmid along with HA-USP10 and/or FLAG-TIA1 plasmid using Lipofectamine 2000. Prepared cell lysates were immunoprecipitated with an anti-HA antibody, and the amounts of Tau, FLAG-TIA1 and HA-USP10 were analyzed by Western blotting using respective antibodies. ( b – e ) HeLa cells were transfected with HA-USP10 plasmid along either with Tau3R1N, Tau4R1N or Tau3R1N/K317M plasmid by lipofection using Lipofectamine 2000. At 48 h after transfection, whole cell lysates ( b , c ), NP-40-soluble fractions and NP-40-insoluble fractions ( d , e ) were prepared and characterized by Western blotting using anti-Tau (TAU-5), anti-phosphorylated Tau (pTau) (AT8), anti-HA, anti-TIA1, anti-α-tubulin and anti-β-actin antibodies.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot

Journal: Scientific Reports

Article Title: USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells

doi: 10.1038/s41598-019-47033-7

Figure Lengend Snippet: The colocalization of USP10 with pTau aggregates in AD brain regions. ( a , b ) Representative immunohistochemical findings of USP10 and pTau in temporal cortex of AD patients (n = 3) are shown. The colocalization of USP10 with pTau aggregates was analyzed by a line profile analysis. Arrows indicate the colocalization suggested by the assay. The relative staining levels of USP10 and pTau on the indicated line were presented ( b ). Lower magnification pictures were presented in Supplementary Fig. . ( c ) The current working model for pathogenic Tau aggregation through SGs and aggresomes.

Article Snippet: Formalin-fixed, paraffin-embedded sections 4-μm-thick were immunostained using a rabbit polyclonal antibody against USP10 (HPA006731; Sigma-Aldrich; diluted 1:2,000, pretreated by heating) and a mouse monoclonal antibody against phosphorylated Tau (AT8; 90206; IGT; diluted 1:200).

Techniques: Immunohistochemical staining, Staining