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rabbit anti psmb5  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc rabbit anti psmb5
    Rabbit Anti Psmb5, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/psmb5/PSMB5+Rabbit+mAb/bio_rxiv__64898__2026__03__28__714855-342-61-65
    Average 93 stars, based on 55 article reviews
    rabbit anti psmb5 - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Western Blot:

    Article Title: Structures of dynamic interactors at native proteasomes by PhIX-MS and cryoelectron microscopy
    Article Snippet: .. Immunoblotting was done by 1 h incubation with primary antibodies; HA-HRP (Cat. No. 2999S; CST), RPN1 (Cat. No. 25430S; CST), PSMB5 (Cat. No. 12919S; CST), ECM29 (Cat. No. AB28666; Abcam), β-actin (Cat. No. AB8226; Abcam) in PierceTM Protein-Free T20 (TBS) Blocking Buffer (Cat. No. 37571; Thermo Fisher Scientific). .. After three 10-min washes in TBST, blots were incubated for 1 h with HRP-conjugated anti-rabbit (Cat. No. 31460; Life Techonologies) or anti-mouse (Cat. No. 31430; Life Techonologies) secondary antibodies, washed again, and developed on a ChemiDoc imaging system (Bio-Rad) using SuperSignal West Pico PLUS substrate (Cat. No. 34577; Thermo Fisher Scientific, Inc).

    Incubation:

    Article Title: Structures of dynamic interactors at native proteasomes by PhIX-MS and cryoelectron microscopy
    Article Snippet: .. Immunoblotting was done by 1 h incubation with primary antibodies; HA-HRP (Cat. No. 2999S; CST), RPN1 (Cat. No. 25430S; CST), PSMB5 (Cat. No. 12919S; CST), ECM29 (Cat. No. AB28666; Abcam), β-actin (Cat. No. AB8226; Abcam) in PierceTM Protein-Free T20 (TBS) Blocking Buffer (Cat. No. 37571; Thermo Fisher Scientific). .. After three 10-min washes in TBST, blots were incubated for 1 h with HRP-conjugated anti-rabbit (Cat. No. 31460; Life Techonologies) or anti-mouse (Cat. No. 31430; Life Techonologies) secondary antibodies, washed again, and developed on a ChemiDoc imaging system (Bio-Rad) using SuperSignal West Pico PLUS substrate (Cat. No. 34577; Thermo Fisher Scientific, Inc).

    Blocking Assay:

    Article Title: Structures of dynamic interactors at native proteasomes by PhIX-MS and cryoelectron microscopy
    Article Snippet: .. Immunoblotting was done by 1 h incubation with primary antibodies; HA-HRP (Cat. No. 2999S; CST), RPN1 (Cat. No. 25430S; CST), PSMB5 (Cat. No. 12919S; CST), ECM29 (Cat. No. AB28666; Abcam), β-actin (Cat. No. AB8226; Abcam) in PierceTM Protein-Free T20 (TBS) Blocking Buffer (Cat. No. 37571; Thermo Fisher Scientific). .. After three 10-min washes in TBST, blots were incubated for 1 h with HRP-conjugated anti-rabbit (Cat. No. 31460; Life Techonologies) or anti-mouse (Cat. No. 31430; Life Techonologies) secondary antibodies, washed again, and developed on a ChemiDoc imaging system (Bio-Rad) using SuperSignal West Pico PLUS substrate (Cat. No. 34577; Thermo Fisher Scientific, Inc).

    Cell-Signaling:

    Article Title: Augment proteasome inhibitor efficacy activates CD8 + T cell-mediated antitumor immunity in breast cancer
    Article Snippet: .. PSMB5 , Cell Signaling Technology , Cat#: 12919; RRID: AB_2798061. .. PSMB6 , Cell Signaling Technology , Cat#: 13267; RRID: AB_2798166.

    Ubiquitin Proteomics:

    Article Title: Xerophenone H, a naturally-derived proteasome inhibitor, triggers apoptosis and paraptosis in lung cancer.
    Article Snippet: Background: Polycyclic polyprenylated acylphloroglucinols (PPAPs) characterized by unique chemical architectures, exhibit diverse pharmacological activities.. Xerophenone H (XeH) is a PPAP extracted from the plant Garcinia multiflora Champ. ex Benth. (Clusiaceae) with a novel and unique chemical structure.. Although in vitro screening has revealed the anti-cancer activity of XeH, whose in vivo effectiveness and mechanistic basis required systematic investigation.

    Article Title: SNPs Give LACTB Oncogene-Like Functions and Prompt Tumor Progression via Dual-Regulating p53.
    Article Snippet: Antibodies against p53 (#ab26), MDM2 p-S186 (ab22710), MDM2 p-S166 (ab170880) and MDM2 (ab16895) were purchased from Abcam. .. Antibodies against GAPDH (#5174S), histone H3 (#4499S), HAtag (#3724S), PSMB5 (#12919S), PSMB6 (#13267S), PSMB7 (#13207S), Ubiquitin (#43124S) and p53 p-S15 (9284S) were purchased from Cell Signalling Technology. .. Secondary antibodies, including goat anti-rabbit IgG H&L (#ab6721), goat anti-mouse IgG H&L (#ab6789), Goat Anti-Mouse IgG H&L (Alexa Fluor 488) (ab150113), Goat Anti-Mouse IgG H&L (Alexa Fluor 647) (ab150115) were purchased from Abcam.

    other:

    Article Title: Suppressing proteasome activity enhances sensitivity to actinomycin D in diffuse anaplastic Wilms tumor.
    Article Snippet: HRP-conjugated secondary antibodies used are Anti-Mouse IgG (Cell Signaling Technology 7076, RRID:AB_330924; 1:10,000) or Anti-Rabbit IgG (Cell Signaling Technology 7074, RRID:AB_2099233; 1:5,000).



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    <t>Proteasome</t> composition detection and hPPSCs-EVs activity. ( A ) Protein expression levels of <t>20S</t> <t>proteasome,</t> PSMD11, and <t>PSMB5.</t> ( B ) Quantitative analysis of 20S <t>proteasome,</t> PSMD11, and PSMB5 protein. ( C ) <t>Proteasome</t> chymotrypsin-like activity assay. n = 3, p < 0.05.
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    <t>Proteasome</t> composition detection and hPPSCs-EVs activity. ( A ) Protein expression levels of <t>20S</t> <t>proteasome,</t> PSMD11, and <t>PSMB5.</t> ( B ) Quantitative analysis of 20S <t>proteasome,</t> PSMD11, and PSMB5 protein. ( C ) <t>Proteasome</t> chymotrypsin-like activity assay. n = 3, p < 0.05.
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    rabbit anti psmb5  (Cell Signaling Technology Inc)
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    <t>Proteasome</t> composition detection and hPPSCs-EVs activity. ( A ) Protein expression levels of <t>20S</t> <t>proteasome,</t> PSMD11, and <t>PSMB5.</t> ( B ) Quantitative analysis of 20S <t>proteasome,</t> PSMD11, and PSMB5 protein. ( C ) <t>Proteasome</t> chymotrypsin-like activity assay. n = 3, p < 0.05.
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    psmb5  (Cell Signaling Technology Inc)
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    MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, <t>PSMB5,</t> PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.
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    psmb5  (Proteintech)
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    MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, <t>PSMB5,</t> PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.
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    keywords skn 1a nrf1 nfe2l1 proteasome proteostasis psmb5  (Proteostasis Therapeutics)
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    MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, <t>PSMB5,</t> PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.
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    a03418  (Boster Bio)
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    A03418, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti psmb5  (Boster Bio)
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    Changes in the major protein components of the proteasome during hemolysis (A) Western blot analysis of proteasomal subunits (PSME1/2, <t>PSMB5/6/7)</t> expression in control and hemolysis groups. (B and C) Quantification of proteasomal subunits (PSME1/2, PSMB5/6/7) expression in the RBC membrane (B) and cytoplasm (C) ( n = 3). (D) Representative immunofluorescence images of proteasomal subunits (PSME1/2, PSMB5/6/7) of hemolytic RBCs ( n = 6). Scale bars = 10 μm. (E) Schematic illustration of the animal model of immune hemolysis. (F and G) Immunofluorescence of proteasomal subunits (PSME1/2, PSMB5/6/7) in hemolytic mouse RBCs (F) and AIHA patient RBCs (G) ( n = 6). Scale bars = 10 μm. (H–J) Statistical analysis of caspase-like activity (H), trypsin-like activity (I), and chymotrypsin-like activity (J) of the membrane proteins after hemolysis ( n = 6). Data were analyzed by Student’s t test (two groups) or one-way ANOVA with Tukey’s test (multiple groups) and are expressed as mean ± SEM. ∗ p < 0.05, ∗∗∗ p < 0.001, ns = no significance.
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    Image Search Results


    Proteasome composition detection and hPPSCs-EVs activity. ( A ) Protein expression levels of 20S proteasome, PSMD11, and PSMB5. ( B ) Quantitative analysis of 20S proteasome, PSMD11, and PSMB5 protein. ( C ) Proteasome chymotrypsin-like activity assay. n = 3, p < 0.05.

    Journal: International Journal of Molecular Sciences

    Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

    doi: 10.3390/ijms27157013

    Figure Lengend Snippet: Proteasome composition detection and hPPSCs-EVs activity. ( A ) Protein expression levels of 20S proteasome, PSMD11, and PSMB5. ( B ) Quantitative analysis of 20S proteasome, PSMD11, and PSMB5 protein. ( C ) Proteasome chymotrypsin-like activity assay. n = 3, p < 0.05.

    Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

    Techniques: Activity Assay, Expressing

    Proteasomes in hPPSCs-EVs suppress HIF-1α expression and ubiquitinated protein accumulation induced by CoCl 2 . ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; CoCl 2 hUVECs processed with 300 μM CoCl 2 for 72 h; CoCl 2 +human umbilical cord mesenchymal stem cells (hUCMSCs)-EVs hUVECs treated with 300 μM CoCl 2 and 75 μg/mL hUCMSCs-EVs for 72 h; and CoCl 2 +hPPSCs-EVs hUVECs treated with 300 μM CoCl 2 and 75 μg/mL hPPSCs-EVs for 72 h (magnification, 200×; scale, 50 μm). ( B ) Quantitative analysis of the number of adherent endothelial cells, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Journal: International Journal of Molecular Sciences

    Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

    doi: 10.3390/ijms27157013

    Figure Lengend Snippet: Proteasomes in hPPSCs-EVs suppress HIF-1α expression and ubiquitinated protein accumulation induced by CoCl 2 . ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; CoCl 2 hUVECs processed with 300 μM CoCl 2 for 72 h; CoCl 2 +human umbilical cord mesenchymal stem cells (hUCMSCs)-EVs hUVECs treated with 300 μM CoCl 2 and 75 μg/mL hUCMSCs-EVs for 72 h; and CoCl 2 +hPPSCs-EVs hUVECs treated with 300 μM CoCl 2 and 75 μg/mL hPPSCs-EVs for 72 h (magnification, 200×; scale, 50 μm). ( B ) Quantitative analysis of the number of adherent endothelial cells, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

    Techniques: Expressing, Control, Cell Culture

    Proteasomes in hPPSCs-EVs inhibit MG-132-induced HIF-1α expression. ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; MG-132 hUVECs subjected to 0.25 μM MG-132 for 72 h; MG-132+hUCMSCs-EVs hUVECs treated with 0.25 μM MG-132 and 75 μg/mL hUCMSCs-EVs for 72 h; and MG-132+hPPSCs-EVs hUVECs treated with 0.25 μM MG-132 and 75 μg/mL hPPSCs-EVs for 72 h (magnification, 200×; scale, 50 μm). ( B ) Quantitative analysis of the number of adherent endothelial cells, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Journal: International Journal of Molecular Sciences

    Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

    doi: 10.3390/ijms27157013

    Figure Lengend Snippet: Proteasomes in hPPSCs-EVs inhibit MG-132-induced HIF-1α expression. ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; MG-132 hUVECs subjected to 0.25 μM MG-132 for 72 h; MG-132+hUCMSCs-EVs hUVECs treated with 0.25 μM MG-132 and 75 μg/mL hUCMSCs-EVs for 72 h; and MG-132+hPPSCs-EVs hUVECs treated with 0.25 μM MG-132 and 75 μg/mL hPPSCs-EVs for 72 h (magnification, 200×; scale, 50 μm). ( B ) Quantitative analysis of the number of adherent endothelial cells, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

    Techniques: Expressing, Control, Cell Culture

    The hPPSCs-EVs proteasomes degrade HIF-1α induced by both CoCl 2 and MG-132. ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; CoCl 2 +MG-132 hUVECs treated with 300 μM CoCl 2 and 0.25 μM MG-132 for 72 h; CoCl 2 +MG-132+hUCMSCs-EVs hUVECs treated with 300 μM CoCl 2 , 0.25 μM MG-132, and 75 μg/mL hUCMSCs-EVs for 72 h; and CoCl 2 +MG-132+hPPSCs-EVs hUVECs treated with 300 μM CoCl 2 , 0.25 μM MG-132, and 75 μg/mL hPPSCs-EVs for 72 h. (magnification: 200×; scale bar: 50 μm) ( B ) Adherent endothelial cell count quantitation, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Journal: International Journal of Molecular Sciences

    Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

    doi: 10.3390/ijms27157013

    Figure Lengend Snippet: The hPPSCs-EVs proteasomes degrade HIF-1α induced by both CoCl 2 and MG-132. ( A ) Representative micrographs of hUVECs in different treatment groups. Groups were set as follows: control hUVECs cultured in serum-free medium; CoCl 2 +MG-132 hUVECs treated with 300 μM CoCl 2 and 0.25 μM MG-132 for 72 h; CoCl 2 +MG-132+hUCMSCs-EVs hUVECs treated with 300 μM CoCl 2 , 0.25 μM MG-132, and 75 μg/mL hUCMSCs-EVs for 72 h; and CoCl 2 +MG-132+hPPSCs-EVs hUVECs treated with 300 μM CoCl 2 , 0.25 μM MG-132, and 75 μg/mL hPPSCs-EVs for 72 h. (magnification: 200×; scale bar: 50 μm) ( B ) Adherent endothelial cell count quantitation, n = 3, p < 0.05. ( C ) The protein expression levels of HIF-1α, 20S proteasome, PSMD11, PSMB5, and ubiquitinated proteins. ( D ) Quantitative analysis of HIF-1α, 20S proteasome, PSMD11, and PSMB5 protein. n = 3, p < 0.05.

    Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

    Techniques: Control, Cell Culture, Cell Characterization, Quantitation Assay, Expressing

    MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

    Journal: The Journal of Biological Chemistry

    Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

    doi: 10.1016/j.jbc.2026.111182

    Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

    Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

    Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

    Changes in the major protein components of the proteasome during hemolysis (A) Western blot analysis of proteasomal subunits (PSME1/2, PSMB5/6/7) expression in control and hemolysis groups. (B and C) Quantification of proteasomal subunits (PSME1/2, PSMB5/6/7) expression in the RBC membrane (B) and cytoplasm (C) ( n = 3). (D) Representative immunofluorescence images of proteasomal subunits (PSME1/2, PSMB5/6/7) of hemolytic RBCs ( n = 6). Scale bars = 10 μm. (E) Schematic illustration of the animal model of immune hemolysis. (F and G) Immunofluorescence of proteasomal subunits (PSME1/2, PSMB5/6/7) in hemolytic mouse RBCs (F) and AIHA patient RBCs (G) ( n = 6). Scale bars = 10 μm. (H–J) Statistical analysis of caspase-like activity (H), trypsin-like activity (I), and chymotrypsin-like activity (J) of the membrane proteins after hemolysis ( n = 6). Data were analyzed by Student’s t test (two groups) or one-way ANOVA with Tukey’s test (multiple groups) and are expressed as mean ± SEM. ∗ p < 0.05, ∗∗∗ p < 0.001, ns = no significance.

    Journal: iScience

    Article Title: Ubiquitination and degradation of CD47 enhances macrophage phagocytosis of hemolytic erythrocytes

    doi: 10.1016/j.isci.2025.114499

    Figure Lengend Snippet: Changes in the major protein components of the proteasome during hemolysis (A) Western blot analysis of proteasomal subunits (PSME1/2, PSMB5/6/7) expression in control and hemolysis groups. (B and C) Quantification of proteasomal subunits (PSME1/2, PSMB5/6/7) expression in the RBC membrane (B) and cytoplasm (C) ( n = 3). (D) Representative immunofluorescence images of proteasomal subunits (PSME1/2, PSMB5/6/7) of hemolytic RBCs ( n = 6). Scale bars = 10 μm. (E) Schematic illustration of the animal model of immune hemolysis. (F and G) Immunofluorescence of proteasomal subunits (PSME1/2, PSMB5/6/7) in hemolytic mouse RBCs (F) and AIHA patient RBCs (G) ( n = 6). Scale bars = 10 μm. (H–J) Statistical analysis of caspase-like activity (H), trypsin-like activity (I), and chymotrypsin-like activity (J) of the membrane proteins after hemolysis ( n = 6). Data were analyzed by Student’s t test (two groups) or one-way ANOVA with Tukey’s test (multiple groups) and are expressed as mean ± SEM. ∗ p < 0.05, ∗∗∗ p < 0.001, ns = no significance.

    Article Snippet: The primary antibodies utilized in this procedure were CD47 (Santa Cruz Biotechnology, Cat# sc-12730), MARCH1 (HUABIO Biotechnology, Cat# ER63906), PSME1 (Abcam, Cat# ab186832), PSME2 (Abcam, Cat# ab183727), PSMB5 (BOSTER, Cat# A03418-1), PSMB6 (ABclonal Technology, Cat# A4053), PSMB7 (BOSTER, Cat# A08095-1), and UBQLN1 (Proteintech Group, Cat# 22126-1-AP).

    Techniques: Western Blot, Expressing, Control, Membrane, Immunofluorescence, Animal Model, Activity Assay