ha usp10  (Addgene inc)

Journal: Cell Communication and Signaling : CCS

Article Title: NDR1 mediates PD-L1 deubiquitination to promote prostate cancer immune escape via USP10

doi: 10.1186/s12964-024-01805-5

Figure Lengend Snippet: USP10 binds to PD-L1 and mediates its deubiquitination. A Identification of the number of candidate proteins interacting with NDR1 and PD-L1 in PC3 cells through CO-IP-MS analysis. B Exogenous PD-L1 coimmunoprecipitated with USP10 in 293 T cells. C USP10 was coimmunoprecipitated with PD-L1 in PC3 cells. D IF experiment was conducted to determine the spatial localization of USP10 and PD-L1. E USP10 expression in prostate cancer cells. F Detection of PD-L1 expression level in PC3 cells after concentration gradient increase of USP10 by WB. G Detection of PD-L1 expression in DU145 cells with siUSP10 gene knockdown by WB. H PD-L1 expression levels in DU145 cells were assessed using a concentration gradient of the USP10 inhibitor (Spautin-1) via immunoblot analysis. I PC3/pCMV and PC3/USP10 cells were subjected with CHX (100 μg/mL) at specified intervals to detect PD-L1-degrading protein levels. J The impact of USP10 on PD-L1 ubiquitination in 293 T cells was detected

Article Snippet: Plasmids were purchased from Sino Biological: pCMV3-STK38-Myc (HG12319-NM), pCMV3-PD-L1-His (HG10084-CH), pCMV3-USP10-HA (HG17833-CY).

Techniques: Co-Immunoprecipitation Assay, Expressing, Concentration Assay, Knockdown, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: NDR1 mediates PD-L1 deubiquitination to promote prostate cancer immune escape via USP10

doi: 10.1186/s12964-024-01805-5

Figure Lengend Snippet: NDR1 promotes binding of USP10 and PD-L1 and inhibits PD-L1 ubiquitination degradation. A Exogenous NDR1 coimmunoprecipitated with USP10 in 293 T cells. B USP10 was coimmunoprecipitated with NDR1 in PC3 cells. C IF experiment revealed the spatial localization of USP10 and NDR1. D Sequential IP assay for NDR1-USP10-PD-L1 interaction. E - F NDR1 knockdown in DU145 cells was assessed for its impact on the protein and mRNA expression of NDR1 and USP10. G-H co-IP assays and IF analysis to explore the relationship between USP10 and PD-L1 in 293T cells, with or without NDR1 overexpression. I: Intranuclear USP10 and PD-L1 expression levels in PC3 cells following a concentration gradient increase of NDR1. J The ubiquitination of intranuclear PD-L1 was examined in 293T cell by WB. K The expression of USP10,NDR1and PD-L1 was detected in NDR1 knockdown or overexpressed DU145 cells with or without Transfected USP10

Article Snippet: Plasmids were purchased from Sino Biological: pCMV3-STK38-Myc (HG12319-NM), pCMV3-PD-L1-His (HG10084-CH), pCMV3-USP10-HA (HG17833-CY).

Techniques: Binding Assay, Knockdown, Expressing, Co-Immunoprecipitation Assay, Over Expression, Concentration Assay, Transfection