primary antibodies against usp22  (Proteintech)

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: USP22 expression is elevated in HF models. ( A ) USP22 expression in datasets GSE116250 and GSE57345 ; HF cell model in vitro was induced by Ang II, ( B ) CCK-8 assay assessing H9c2 cell viability; ( C ) TUNEL assay detecting apoptosis levels in H9c2 cells; ( D , E ) USP22 expression in H9c2 cells measured by RT-qPCR ( D ) and WB ( E ); CHF rat model was established through AAC, ( F ) Hemodynamic assessment of cardiac function in AAC-induced rats; ( G ) Biochemical analysis of serum ANP, cTnT, and CK-MB levels in rats. ( H , I ) USP22 expression in rat cardiac tissues detected by RT-qPCR ( H ) and IHC ( I ). Comparisons between two groups were analyzed using the t -test. * P < 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Expressing, In Vitro, CCK-8 Assay, TUNEL Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: USP22 knockdown alleviates cardiomyocyte injury by inhibiting HIF-1α protein stability. USP22 knockdown was mediated by shRNA in H9c2 cells, ( A , B ) Validation of USP22 knockdown efficiency by RT-qPCR ( A ) and WB ( B ); cardiomyocyte injury was induced by Ang II, ( C ) Assessment of H9c2 cell viability using CCK-8 assay; ( D ) Quantification of H9c2 cell apoptosis by TUNEL assay; ( E ) Analysis of apoptosis-related proteins BAX and Bcl-2 expression by WB. Comparisons between two groups were analyzed using the t -test. Multi-group comparisons were analyzed by one-way ANOVA, followed by Tukey’s post-hoc test. * P < 0.05, ns P > 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Knockdown, shRNA, Biomarker Discovery, Quantitative RT-PCR, CCK-8 Assay, TUNEL Assay, Expressing

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: USP22 knockdown alleviates myocardial injury in CHF rats. Cardiac-specific knockdown of USP22 was performed in rats subjected to AAC-induced CHF, ( A , B ) USP22 expression in rat cardiac tissues assessed by RT-qPCR ( A ) and IHC ( B ); ( C ) Hemodynamic parameters evaluating cardiac function; ( D ) Biochemical quantification of serum ANP, cTnT, and CK-MB levels in rats. ( E , F ) Representative images of H&E ( E ) and Masson’s trichrome ( F ) staining evaluating cardiomyocyte damage and fibrosis, respectively. Comparisons between two groups were analyzed using the t -test. * P < 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Staining

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: HIF-1α overexpression rescues cardiomyocytes from the protective effects of USP22 knockdown. ( A , B ) Impact of USP22 knockdown on HIF-1α mRNA (A; RT-qPCR) and protein ( B ; WB) expression under Ang II induction; ( C ) Effect of USP22 knockdown on HIF-1α protein stability assessed by CHX chase assay; ( D ) USP22-HIF-1α interaction verified by Co-IP; ( E ) Ubiquitination analysis evaluating the effect of USP22 knockdown on HIF-1α polyubiquitination levels using HA-Ub overexpression and Co-IP; HIF-1α was overexpressed in the context of USP22 knockdow, ( F , G ) Validation of HIF-1α overexpression efficiency by RT-qPCR ( F ) and WB ( G ) in USP22-knockdown cells; ( H ) H9c2 cell viability assessed by CCK-8 assay; ( I ) Quantification of H9c2 cell apoptosis by TUNEL assay; ( J ) Expression of apoptosis-related proteins BAX and Bcl-2 analyzed by WB. Comparisons between two groups were analyzed using the t -test, while comparisons across multiple groups were performed with one-way or two-way ANOVA followed by Tukey’s post hoc test. * P < 0.05, ns P > 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Over Expression, Knockdown, Quantitative RT-PCR, Expressing, Co-Immunoprecipitation Assay, Ubiquitin Proteomics, Biomarker Discovery, CCK-8 Assay, TUNEL Assay

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: PATZ1 promotes USP22 transcription. ( A ) Prediction of upstream transcriptional regulators of USP22 using the JASPAR database; ( B ) Intersection of predicted USP22 regulators with DEGs from GSE116250 and GSE57345 HF datasets; ( C ) Correlation analysis of ZNF460, PATZ1, and MAZ expression with USP22 levels in left ventricular tissue using GEPIA 2.0; ( D ) PATZ1 expression in cardiac tissues from HF patients within datasets GSE116250 and GSE57345 ; ( E , F ) PATZ1 expression in Ang II-treated H9c2 cells assessed by RT-qPCR ( E ) and WB ( F ); Following shRNA-mediated PATZ1 knockdown, ( G , H ) expression levels of PATZ1 and USP22 measured by RT-qPCR ( G ) and WB ( H ); ( I ) Binding of PATZ1 to the USP22 promoter region confirmed by ChIP-qPCR; ( J ) Impact of PATZ1 knockdown on USP22 promoter activity assessed by dual-luciferase reporter assay. Comparisons between two groups were analyzed using the t -test. * P < 0.05, ns P > 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, shRNA, Knockdown, Binding Assay, ChIP-qPCR, Activity Assay, Luciferase, Reporter Assay

Journal: Scientific Reports

Article Title: Knockdown of PATZ1 alleviates chronic heart failure through the USP22/HIF-1α axis

doi: 10.1038/s41598-025-24336-6

Figure Lengend Snippet: PATZ1 knockdown alleviates HF via the USP22/HIF-1α axis. H9c2 cells were subjected to PATZ1 knockdown with or without USP22 overexpression, ( A , B ) PATZ1 and USP22 expression levels in H9c2 cells assessed by RT-qPCR (A) and WB ( B ); HF cell model in vitro was induced by Ang II, ( C ) H9c2 cell viability measured by CCK-8 assay; ( D ) Quantification of H9c2 apoptosis by TUNEL assay; ( E ) Expression of PATZ1, apoptosis-related proteins (BAX, Bcl-2) and HIF-1α analyzed by WB; AAC-induced CHF rats were subjected to AAV-sh-PATZ1 with or without USP22 overexpression, ( F ) IHC detection of PATZ1, USP22, and HIF-1α expression in cardiac tissues; ( G ) Hemodynamic assessment of cardiac function in rats; ( H ) Biochemical analysis of serum ANP, cTnT, and CK-MB levels in rat serum; ( I , J ) Representative images of H&E ( I ) and Masson’s trichrome ( J ) staining evaluating myocardial histoarchitecture and fibrosis, respectively. Comparisons across multiple groups were performed with one-way ANOVA followed by Tukey’s post hoc test. * P < 0.05, ns P > 0.05.

Article Snippet: Primary antibodies against USP22 (1:200, 55110-1-AP, Proteintech), HIF-1α (1:200, ab179483, Abcam), and PATZ1 (1:200, Cat# SAB2101726, Sigma-Aldrich) were applied and sections were incubated overnight at 4°C.

Techniques: Knockdown, Over Expression, Expressing, Quantitative RT-PCR, In Vitro, CCK-8 Assay, TUNEL Assay, Staining

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 mRNA and protein levels in GC tissues and cell lines. ( A ) Histogram plots show USP22 mRNA levels based on qRT-PCR analysis in 40 pairs of GC (C) and adjacent noncancerous (N) tissues. GAPDH was used as an endogenous control. ( B ) Representative images show western blot analysis of USP22 protein levels in eight pairs of GC ( C ) and adjacent noncancerous tissues (N). β-actin was used as endogenous control. ( C – D ) The levels of USP22 mRNA ( C ) and protein ( D ) in GC cell lines and normal gastric mucosal epithelial cells are shown in the histogram plots and representative images, respectively. GAPDH and β-actin were used as the controls in the qRT-PCR and western blot analysis, respectively. Data are expressed as mean ± SD of three replicates. * p < 0.05 is considered statistically significant.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: Quantitative RT-PCR, Control, Western Blot

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 silencing inhibits in vitro GC cell proliferation. ( A ) Representative images show USP22 expression in BGC-823 and HGC-27 cells that were transfected with 100 nM siNC, siUSP22-1, or siUSP22-2 cultured for 24 h. Untransfected (mock) GC cells were used as controls. β-actin was used as an endogenous control. ( B ) MTT assay results show viability of siNC or siUSP22-1 transfected GC cells at days 1, 2, 3, and 4. ( C ) Flow cytometry analysis shows the effects of USP22 knockdown in GC cells. The percentage of G1, S-, and G2M cells in siNC or siUSP22-1 transfected GC cells are shown in the histogram plots. ( D ) EdU assay results show cell proliferation status of siNC or siUSP22-1 transfected GC cells. The plots show the percentage of EdU-positive cells in various samples. Magnification: 100×. ( E ) Histograms (right) show the total number of colonies in BGC-823 and HGC-27 cells that are uninfected (Mock) or infected with lentiviruses carrying pLKO.1 or pLKO.1-shUSP22 vectors. Representative images of colony formation assay are also shown. Note: Data are expressed as mean ± SD of three replicates; * p < 0.05; ** p < 0.01 compared with mock or siNC group in ( B – D ); * p < 0.05 compared with the mock or pLKO.1 group in ( E ).

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: In Vitro, Expressing, Transfection, Cell Culture, Control, MTT Assay, Flow Cytometry, Knockdown, EdU Assay, Infection, Colony Assay

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 knockdown induces in vitro GC cell apoptosis. The results of ( A ) Annexin V/PI staining, ( B ) TUNEL, and ( C ) caspase-3 activity assays in BGC-823 and HGC-27 cells that are untransfected (Mock) or transfected with 100 nM siNC or siUSP22-1are shown. Note: Data are expressed as mean ± SD of three replicates; * p < 0.05 compared with the mock or siNC group.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: Knockdown, In Vitro, Staining, TUNEL Assay, Activity Assay, Transfection

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 silencing suppresses in vitro GC cell migration and invasion. ( A – B ) Transwell assay results show total number of ( A ) migratory and ( B ) invasive BGC-823 and HGC-27 cells that are untransfected (Mock) or transfected with 100 nM siNC or siUSP22-1 at 48 h after transfection. Note: Magnification: 200×; Data are expressed as mean ± SD of three replicates. * p < 0.05 compared with mock or siNC group.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: In Vitro, Migration, Transwell Assay, Transfection

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 promotes in vitro GC progression via c-Myc/NAMPT/ SIRT1-dependent FOXO1 and YAP signaling pathways. ( A ) Representative images show the levels of USP22, c-Myc, NAMPT, and SIRT1 proteins in BGC-823 and HGC-27 cells transfected with 100 nM siNC or siUSP22-1. The cells were analyzed at 24 h after transfection. β-actin was used as an endogenous control. ( B ) ChIP assay analysis shows binding efficiency of c-Myc in the promoter region of NAMPT in BGC-823 and HGC-27 cells that were transfected with 100 nM sic-Myc or siNC. ( C ) Representative images show the levels of c-Myc and NAMPT proteins in BGC-823 and HGC-27 cells that were transfected with 100 nM sic-Myc or siNC. β-actin was used as endogenous control. ( D ) ChIP assay analysis shows binding efficiency of NAMPT in the promoter region of SIRT1 in BGC-823 and HGC-27 cells that were transfected with 100 nM siNAMPT or siNC. ( E ) Representative images show the levels of NAMPT and SIRT1 protein in BGC-823 and HGC-27 cells were transfected with 100 nM siNAMPT or siNC. β-actin was used as endogenous control. ( F – G ) Representative images show the levels of ( F ) Sirt1, FOXO1, Ki67, Cyclin D1, Bax, and Bcl-2, and ( G ) Sirt1, YAP, MMP-2, and MMP-9 proteins in BGC-823 and HGC-27 cells transfected with 100 nM siSirt1 or siNC, or cells treated with 100 μM EX527, a Sirt1 inhibitor. β-actin was used as an endogenous control.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: In Vitro, Protein-Protein interactions, Transfection, Control, Binding Assay

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 depletion reduces in vivo GC growth and metastasis. Six-week-old male SCID mice were subcutaneously injected with stably-expressed pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells into the hind flanks or through the tail vein. ( A ) Histogram shows tumor volume at weeks 1, 2, 3 and 4 after subcutaneous injection of SCID mice (n=5 each) with BGC-823-luc cells stably-expressing pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells into the hind flanks at 1, 2, 3 and 4 weeks. ( B ) Analysis of tumor growth progression by in vivo luciferase imaging of the xenografts at day 21 after subcutaneous injection of SCID mice (n=5 each) with stably-expressed pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells. ( C ) Representative photographs of xenograft tumors at day 42 from SCID mice subcutaneously injected with stably-expressed pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells. ( D ) TUNEL assay results show the total number of apoptotic cells in the xenograft tumor sections derived at day 42 from from SCID mice subcutaneously injected with stably-expressed pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells. ( E ) Representative images and ( F ) Total number of s metastatic nodules in the lungs at day 28 from SCID mice that were injected with stably-expressed pLKO.1-shUSP22-1 or pLKO.1 BGC-823-luc cells. Note: Data are expressed as mean ± SD of three replicates; * p < 0.05, ** p < 0.01 compared with the mock or pLKO.1 group.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: In Vivo, Injection, Stable Transfection, Expressing, Luciferase, Imaging, TUNEL Assay, Derivative Assay

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: The correlation of USP22 expression with clinicopathological features of gastric cancer.

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Oncogenic USP22 supports gastric cancer growth and metastasis by activating c-Myc/NAMPT/SIRT1-dependent FOXO1 and YAP signaling

doi: 10.18632/aging.102410

Figure Lengend Snippet: USP22 overexpression positively correlates with advanced histological grade, metastasis, and poor prognosis in GC patients. ( A ) Representative images of IHC staining of USP22 and ( B ) USP22 expression scores in the sections of non-cancerous and GC tissues at various stages of differentiation. Scale bar: 10 μm. ( C ) USP22 expression scores in metastatic and non-metastatic GC tissue samples (n = 40). ( D ) Relationship between levels of USP22 expression based on IHC and the five-year overall survival (OS) rates after surgery in GC patients with high-USP22 (n = 107) and low-USP22 (n = 79) expression. Note: Data are expressed as mean ± SD of three replicates; * p < 0.05 compared with non-cancerous tissues or well-differentiated tissues in ( B ); * p < 0.05 compared with non-metastatic GC tissues in ( C ); # p < 0.05 compared with moderate-differentiated tissues in ( B ).

Article Snippet: Then, the sections were incubated with primary antibody against USP22 (1:150 dilution, Cell Signaling Technology) at 4°C overnight followed by incubation for 20 min with the biotinylated secondary antibody using the ChemMate EnVision Kit (DAKO, Hamburg, Germany).

Techniques: Over Expression, Immunohistochemistry, Expressing