rabbit monoclonal antibodies against impdh2  (Servicebio Inc)

Journal: Cell Death & Disease

Article Title: USP5 regulates purine metabolism and represents a therapeutic target in esophageal cancer

doi: 10.1038/s41419-026-08683-4

Figure Lengend Snippet: A MS analysis identified proteins commonly enriched in KYSE450 and KYSE30 cells. B , C Endogenous and exogenous co-IP confirmed the interaction between USP5 and IMPDH2. D Molecular docking of USP5 with IMPDH2. E IMPDH2 protein levels were assessed by WB after USP5 knockdown. F IMPDH2 protein stability was assessed by WB after USP5 overexpression, and CHX treatment at various time points. G KYSE150 cells were treated with 3-MA or MG132 after USP5 knockdown, and IMPDH2 stability was analyzed by WB after CHX treatment. H Flag-USP5 WT or Flag-USP5 C335A plasmids were transfected into HEK293 cells, and IMPDH2 ubiquitination was assessed by Western blot after 6 h of MG132 treatment. I In HEK293 cells, Myc-IMPDH2, Myc-IMPDH2-K109R, Myc-IMPDH2-K124R, Myc-IMPDH2-K167R, Myc-IMPDH2-K375R, Myc-IMPDH2-K438R, Myc-IMPDH2-K489R plasmids were transfected, followed by MG132 treatment, and IMPDH2 ubiquitination levels were assessed by Western blot. J Endogenous and exogenous co-IP with SYVN1 and IMPDH2 antibodies was performed by WB in KYSE150 and KYSE450 cells. K SYVN1 plasmid was transfected into HEK293 cells, followed by MG132 treatment, and IMPDH2 ubiquitination levels were assessed by Western blot.

Article Snippet: Primary antibodies included rabbit monoclonal antibodies against IMPDH2 and USP5, and a murine monoclonal antibody against Ki67 (1:400; GB121141 ; Servicebio Technology).

Techniques: Co-Immunoprecipitation Assay, Knockdown, Over Expression, Transfection, Ubiquitin Proteomics, Western Blot, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: USP5 regulates purine metabolism and represents a therapeutic target in esophageal cancer

doi: 10.1038/s41419-026-08683-4

Figure Lengend Snippet: A Measurement of guanine levels in KYSE410 cells following IMPDH2 overexpression. B Measurement of guanine levels in KYSE450 and KYSE150 cells after IMPDH2 knockdown. C Guanine levels in KYSE450 cells after treatment with MPA (mycophenolic acid). D , E MTT and foci formation assays in KYSE150 and KYSE450 cells treated with MPA and supplemented with exogenous guanine. F MTT assay was conducted to measure the proliferation in KYSE150 and KYSE450 cells after IMPDH2 knockdown with exogenous guanine supplementation. G – I CDX model combining IMPDH2 knockdown and free-purine diet; tumor weight ( H ) and volume ( I ) measurements. J In KYSE150 and KYSE450 cells with IMPDH2 knockdown, exogenous guanine was added, and guanine levels were measured. K Western blot validation of USP5 knockdown and IMPDH2 overexpression in KYSE70 cells. L MTT assay evaluating the effect of USP5 knockdown and IMPDH2 overexpression on cell proliferation in KYSE70 cells. M – P . Foci formation and soft agar assays assessing the impact of USP5 knockdown and IMPDH2 overexpression on clonogenic capacity of ESCC cells. Q In USP5-knockdown KYSE150 and KYSE450 cells, Myc-IMPDH2 plasmid was transfected, and guanine levels were measured. R In USP5-knockdown KYSE150 and KYSE450 cells, Flag-USP5, Myc-IMPDH2, and Myc-IMPDH2-K489R plasmids were transfected, followed by MG132 treatment, and guanine levels were measured. S In KYSE150 and KYSE450 cells, Flag-SYVN1 and Myc-IMPDH2 plasmids were transfected, followed by MG132 treatment, and guanine levels were measured. Data are presented as mean ± SD. Statistical significance was assessed by unpaired two-tailed Student’s t test ( A , C , R , S ) two-way ANOVA ( L ) or one-way ANOVA ( B , D , E , F , H , I , J , O , P , Q ) as appropriate. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies included rabbit monoclonal antibodies against IMPDH2 and USP5, and a murine monoclonal antibody against Ki67 (1:400; GB121141 ; Servicebio Technology).

Techniques: Over Expression, Knockdown, MTT Assay, Western Blot, Biomarker Discovery, Plasmid Preparation, Transfection, Two Tailed Test

Journal: Cell Death & Disease

Article Title: USP5 regulates purine metabolism and represents a therapeutic target in esophageal cancer

doi: 10.1038/s41419-026-08683-4

Figure Lengend Snippet: A Western blot analysis of USP5 and IMPDH2 protein expression after MDZ treatment. MDZ represents Mebendazole. B Guanine levels were measured in KYSE150 and KYSE450 cells after 48 hours of MDZ treatment. C USP5 protein stability was assessed in KYSE150 and KYSE450 cells after MDZ treatment followed by CHX treatment. D In KYSE70 and KYSE30 cells, MDZ treatment followed by MG132 treatment was performed, and IMPDH2 ubiquitination levels were assessed by Western blot. E In KYSE450 and KYSE30 cells, SYVN1 was transfected, followed by MDZ (Mebendazole) treatment and MG132 treatment, and IMPDH2 ubiquitination levels were assessed by Western blot. F Immunofluorescence staining was performed to detect USP5 protein levels after Mebendazole treatment. G MTT assay was performed to assess cell proliferation levels of KYSE30, KYSE450, KYSE150, and KYSE510 cells after MDZ treatment. H , I Foci formation and soft agar assays were performed to assess the colony formation ability of KYSE30, KYSE450, KYSE150, and KYSE510 cells after MDZ treatment. J Tumor images from the PDX model after intragastric administration of MDZ. K , L Tumor weight and tumor volume after MDZ (Mebendazole) treatment. Data are presented as mean ± SD. Statistical significance was assessed by two-way ANOVA ( G , K ) or one-way ANOVA ( B , H , I , L ) as appropriate. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies included rabbit monoclonal antibodies against IMPDH2 and USP5, and a murine monoclonal antibody against Ki67 (1:400; GB121141 ; Servicebio Technology).

Techniques: Western Blot, Expressing, Ubiquitin Proteomics, Transfection, Immunofluorescence, Staining, MTT Assay

Journal: Cell Death & Disease

Article Title: USP5 regulates purine metabolism and represents a therapeutic target in esophageal cancer

doi: 10.1038/s41419-026-08683-4

Figure Lengend Snippet: A MTT assay was performed to assess the proliferation of KYSE150 and KYSE450 cells after combined treatment with MDZ and Oxa. Oxa represents Oxaliplatin. B The sensitivity to Oxa was assessed in KYSE150 and KYSE450 cells after USP5 or IMPDH2 knockdown. C . MTT assay was assessed in KYSE450 and KYSE510 cells treated with a combination of MDZ and Oxa. D MTT assay was assessed in KYSE450 and KYSE510 cells treated with a combination of USP5 knockdown and Oxa. E – G The PDX model was used to evaluate the combined treatment of MDZ (25 mg/kg) and Oxa (5 mg/kg) ( E ), with tumor weight ( F ) and tumor volume ( G ) measured and analyzed. H Schematic diagram of the mechanism by which USP5 stabilizes IMPDH2 expression and promotes guanine biosynthesis. The USP5-targeting agent mebendazole enhances oxaliplatin sensitivity when used in combination therapy. Data are presented as mean ± SD. Statistical significance was assessed by two-way ANOVA ( A , G ) or one-way ANOVA ( B , C , D , F ) as appropriate. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies included rabbit monoclonal antibodies against IMPDH2 and USP5, and a murine monoclonal antibody against Ki67 (1:400; GB121141 ; Servicebio Technology).

Techniques: MTT Assay, Knockdown, Expressing