anti fdx1  (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: Communications biology

Article Title: A chromatin-focused CRISPR screen identifies USP22 as a barrier to somatic cell reprogramming.

doi: 10.1038/s42003-025-07899-y

Figure Lengend Snippet: Fig. 3 | USP22 loss results in differentiation defects. A Schematic representation of generating USP22 knockout iPSC clones used in Fig. 3B. B Western blot image showing USP22proteinlevels iniPSCs generatedfromCas9and USP22targetinggRNA expressing fibroblasts. Tubulin was used as a loading control. C Western blot image showing USP22 protein levels in iPSC clones generated from Cas9 and USP22 targeting gRNA expressing

Article Snippet: Boiled samples and protein markers (Bio-Rad, cat. # 161-0374) were loadedontopre-cast SDS-PAGEgels (Bio-Rad, cat. # 456-1084).Using Bio-Rad trans-blot turbo transfer system at mixed weight transfer settings, proteins were transferred onto PVDFmembrane (Bio-Rad, cat. # 1620177), followed by incubation in 5% blotting grade blocker (Bio-Rad, cat. # 1706404) solution for 1 h. Membranes were incubated with antibodies against USP22 (NOVUS, cat. # NBP1-49644), Ubiquityl-Histone H2A (Lys119) (Cell Signaling, cat. # 8240S), Ubiquityl-Histone H2B (Lys120) (Cell Signaling, cat. # 5546S), H2B (Abcam, cat. #ab52599-1:20000), Tubulin and GAPDH (Abcam, cat. #ab9485) at 1:1000 ratio overnight at 4 °C.

Techniques: Knock-Out, Clone Assay, Western Blot, Expressing, Control, Generated

Journal: Communications biology

Article Title: A chromatin-focused CRISPR screen identifies USP22 as a barrier to somatic cell reprogramming.

doi: 10.1038/s42003-025-07899-y

Figure Lengend Snippet: Fig. 4 | USP22 loss stabilizes pluripotency network during reprogramming. A Volcano-plot of RNA-seq data comparing USP22 KO and control cells during reprogramming. B Gene ontology analysis for differentially regulated genes upon USP22 knockout. Top 5 upregulated and downregulated gene sets are shown. C GSEA p-value and normalized enrichment scores on pre-ranked gene lists according to log2FC value for comparisons of reprogramming fibroblasts expressing USP22 gRNA to NT1 for allgene sets available at The Molecular Signatures Database (mSigDB). D GSEA results for pluripotency- and fibroblast-related gene sets upon USP22 loss during reprogramming. E Relative expression levels of pluripotency-

Article Snippet: Boiled samples and protein markers (Bio-Rad, cat. # 161-0374) were loadedontopre-cast SDS-PAGEgels (Bio-Rad, cat. # 456-1084).Using Bio-Rad trans-blot turbo transfer system at mixed weight transfer settings, proteins were transferred onto PVDFmembrane (Bio-Rad, cat. # 1620177), followed by incubation in 5% blotting grade blocker (Bio-Rad, cat. # 1706404) solution for 1 h. Membranes were incubated with antibodies against USP22 (NOVUS, cat. # NBP1-49644), Ubiquityl-Histone H2A (Lys119) (Cell Signaling, cat. # 8240S), Ubiquityl-Histone H2B (Lys120) (Cell Signaling, cat. # 5546S), H2B (Abcam, cat. #ab52599-1:20000), Tubulin and GAPDH (Abcam, cat. #ab9485) at 1:1000 ratio overnight at 4 °C.

Techniques: RNA Sequencing, Control, Knock-Out, Expressing