usp10  (Proteintech)

Journal: International Journal of Biological Sciences

Article Title: Co-targeting MRPS7-23 synergistically enhances cisplatin efficacy to suppress nasopharyngeal carcinoma growth and metastasis

doi: 10.7150/ijbs.115523

Figure Lengend Snippet: USP10 depletion reduces expression of MRPS7 and MRPS23 and suppresses xenograft tumor growth. (A-C) The protein of USP10 co-immunoprecipitated with MRPS7 and MRPS23 was verified by mass spectrometry. (D) The co-localization of MRPS7 (green), MRPS23 (red) and USP10 (purple) detected by immunofluorescence assay in NPC cells. The scale bar is 10 μm. (E) The binding interface between MRPS7 (yellow) and USP10 (purple) was predicted using computational molecular docking. (F) The binding interface between MRPS23 (yellow) and USP10 (purple) was predicted using computational molecular docking. (G) Direct interaction between USP10 and MRPS7 measured by surface plasmon resonance. (H) Direct interaction between USP10 and MRPS23 measured by surface plasmon resonance. (I) OS curves of low and high expressed of USP10. (J) Bioinformatic analysis of TCGA data revealed differential expression of USP10 in HNSC tumors compared to normal tissues. (K) Western blot analysis of MRPS7, MRPS23 and USP10 protein levels in NPC cells with or without knockdown of USP10 after the treatment with MG132. (L) Following co-transfection with HA-Ub-K48O and either an empty vector or Flag-MRPS7/23 plasmids, NPC cells were treated with MG132 and lysates were immunoprecipitated under denaturing conditions with the specified antibodies. (M) Western blot analysis was conducted to assess MRPS7, MRPS23, and β-catenin expression in USP10-knockdown NPC cells, with or without concomitant overexpression of MRPS7 and MRPS23. (N) The photo of excised Cne2 tumors. (O) Tumor volumetric measurements were evaluated at predetermined intervals. (P) The measurement of tumor weight in excised Cne2 tumors. (Q-R) Ki67 staining and expression of tumor tissue. * p < 0.05 and *** p < 0.001.

Article Snippet: For western blot analysis, After SDS-PAGE (10%) and PVDF transfer (Millipore), membranes were blocked (5% milk, 1 h RT) then incubated with primary antibodies (4°C, overnight): MRPS7 (1:1000, Abcam, UK), MRPS23 (1:1000, Proteintech, USA), β-catenin (1:1000, Proteintech, USA), Slug (1:1000, Proteintech, USA), Vimentin (1:1000, Proteintech, USA), E-cadherin (1:1000, Proteintech, USA), USP10 (1:1000, Proteintech, USA), SOX2 (1:1000, Proteintech, USA), Oct4 (1:1000, Abcam, UK), and Nanog (1:1000, Abcam, UK).

Techniques: Expressing, Immunoprecipitation, Mass Spectrometry, Immunofluorescence, Binding Assay, SPR Assay, Quantitative Proteomics, Western Blot, Knockdown, Cotransfection, Plasmid Preparation, Over Expression, Staining

Journal: International Journal of Biological Sciences

Article Title: Co-targeting MRPS7-23 synergistically enhances cisplatin efficacy to suppress nasopharyngeal carcinoma growth and metastasis

doi: 10.7150/ijbs.115523

Figure Lengend Snippet: Therapeutic targeting of USP10 by spautin-1 potentiates cisplatin efficacy to suppress tumor growth and metastasis in NPC. (A-B) Cne2 and C666 cells were incubated with Spautin-1, cisplatin, or combination therapy at graded concentrations for 48 h, with viability determined by CCK-8 assay. (C-D) Synergistic effects of Spautin-1/Cisplatin combination in Cne2 and C666 cells were quantitatively analyzed using CalcuSyn software. (E-F) Dose-dependent combination indexes (CIs) and fraction affected (Fa) values for Spautin-1/Cisplatin treatment in both cell lines are presented. (G-I) The Spautin-1/Cisplatin combination significantly suppressed the migratory capacity of NPC cells. (J) Images of resected Cne2 xenograft tumors. (K) Tumor growth kinetics were monitored at specified time points. (L) Tumor weight of excised Cne2 tumors. (M) The mice were weighted on the indicated days. (N) Ki67, slug, E-cadherin and SOX2 staining and expression of tumor tissue. (O) The relative expression of Ki67, slug, E-cadherin and SOX2 in tumor tissue. (P) Schematic diagram of spautin-1 and cisplatin treatment on NPC lung metastasis. (Q-R) H&E-stained lung tissue of lung metastasis model, with subsequent quantification of microscopically detectable metastatic foci. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: For western blot analysis, After SDS-PAGE (10%) and PVDF transfer (Millipore), membranes were blocked (5% milk, 1 h RT) then incubated with primary antibodies (4°C, overnight): MRPS7 (1:1000, Abcam, UK), MRPS23 (1:1000, Proteintech, USA), β-catenin (1:1000, Proteintech, USA), Slug (1:1000, Proteintech, USA), Vimentin (1:1000, Proteintech, USA), E-cadherin (1:1000, Proteintech, USA), USP10 (1:1000, Proteintech, USA), SOX2 (1:1000, Proteintech, USA), Oct4 (1:1000, Abcam, UK), and Nanog (1:1000, Abcam, UK).

Techniques: Incubation, CCK-8 Assay, Software, Staining, Expressing

Journal: International Journal of Biological Sciences

Article Title: Co-targeting MRPS7-23 synergistically enhances cisplatin efficacy to suppress nasopharyngeal carcinoma growth and metastasis

doi: 10.7150/ijbs.115523

Figure Lengend Snippet: MRPS7, MRPS23, and USP10 expression predicts poor outcomes and chemoresistance in nasopharyngeal carcinoma. (A-C) Immunohistochemical (IHC) staining showing different intensities of MRPS7, MRPS23, and USP10 expression in NPC tissues. Scale bar, 100 μm. (D-E) Comparative IHC analysis of MRPS7, MRPS23, and USP10 expression in tumor versus adjacent non-tumor tissues (n = 41). Scale bar, 100 μm. (F) IHC images of MRPS7, MRPS23, and USP10 in NPC patient samples. Scale bar, 100 μm. (G) Correlation analysis of MRPS7 with MRPS23, MRPS7 with USP10, and MRPS23 with USP10 in NPC tissues (n = 41). (H-I) Kaplan-Meier survival curves demonstrating that high expression of MRPS7, MRPS23, and USP10 is associated with shorter disease-free survival. (J) MRPS7, MRPS23 and USP10 expression, as evaluated by IHC staining, was associated with the response to cisplatin treatment.

Article Snippet: For western blot analysis, After SDS-PAGE (10%) and PVDF transfer (Millipore), membranes were blocked (5% milk, 1 h RT) then incubated with primary antibodies (4°C, overnight): MRPS7 (1:1000, Abcam, UK), MRPS23 (1:1000, Proteintech, USA), β-catenin (1:1000, Proteintech, USA), Slug (1:1000, Proteintech, USA), Vimentin (1:1000, Proteintech, USA), E-cadherin (1:1000, Proteintech, USA), USP10 (1:1000, Proteintech, USA), SOX2 (1:1000, Proteintech, USA), Oct4 (1:1000, Abcam, UK), and Nanog (1:1000, Abcam, UK).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry