human stat1 cdna (Addgene inc)
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Addgene inc
human stat1 cdna
Human Stat1 Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+stat1+cdna/pmc12711663-531-10-18?v=Addgene+inc
Average 93 stars, based on 6 article reviews
Human Stat1 Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+stat1+cdna/pmc12711663-531-10-18?v=Addgene+inc
Average 93 stars, based on 6 article reviews
human stat1 cdna - by Bioz Stars,
2026-07
93/100 stars
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1) Product Images from "Therapeutic stress triggers tumor STAT1 acetylation to disarm immunotherapy"
Article Title: Therapeutic stress triggers tumor STAT1 acetylation to disarm immunotherapy
Journal: Cell Reports Medicine
doi: 10.1016/j.xcrm.2025.102448
Figure Legend Snippet: STAT1 Lys637 acetylation correlates with poor response to ICB therapy (A) Schematic of a syngeneic murine oral cancer model receiving anti-PD1 injection. The murine oral squamous cell carcinoma cell line MOC-L2-1 with Stat1 knockdown (shmStat1) and reconstituted with human STAT1 (hSTAT1(WT) or hSTAT1(K637Q) or hSTAT1(K637R)) was inoculated subcutaneously into C57BL/6J mice until tumors reached a volume of 100 mm 3 . Five doses of anti-PD1 or isotype IgG were administered to tumor-bearing mice. n = 9–10 per group. (B) Tumor growth inhibition (TGI, %) calculated as the relative change in tumor volume between day 0 and day 38 in different groups. Data presented as mean ± SEM. ∗∗∗ p < 0.001. (C) Tumor weight in the mouse experiments. Data presented as mean ± SEM. ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, not significant. (D) Kaplan-Meier overall survival curves for HNSCC patients ( n = 63) stratified by H-score cutoff of 166 with median follow-up of 8.0 months (range 0.5–45.1). (E) Kaplan-Meier overall survival curves for GC patients ( n = 46) stratified by H-score cutoff of 166 with median follow-up of 9.7 months (range 1.97–60.2). (F) Kaplan-Meier overall survival curves for hepatocellular carcinoma (HCC) patients ( n = 39) stratified by H-score cutoff of 166 with median follow-up of 15.5 months (range 3.1–81.1). (G) Comparison of STAT1 Lys637 acetylation levels between HNSCC responders ( n = 27) and non-responders ( n = 36) to ICB treatment. Statistical analyses were performed using an unpaired Student’s t test. ∗∗ p < 0.01. (H) Comparison of STAT1 K637 acetylation levels between HNSCC disease control patients ( n = 42) and those with progressive disease ( n = 21) following ICB therapy. Statistical analyses were performed using an unpaired Student’s t test. ∗∗∗ p < 0.001. See also , , , , and .
Techniques Used: Injection, Knockdown, Inhibition, Comparison, Control
Figure Legend Snippet: Impaired IFN-γ response and reduced STAT1 protein in cetuximab-resistant HNSCC (A) RT-qPCR of IFN-γ response-associated gene expression, including tumor immunology-related genes (upper), antiviral-related genes (middle), and antigen processing and presentation genes (lower) in OECM-1-WT and OECM-1-Ctx R cells. n = 3 (each with two technical replicates). The cells were then treated with IFN-γ (100 ng/mL) for 24 h. Data are presented as mean ± SD. Statistical significance was determined using unpaired Student’s t test. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, not significant. (B) RT-qPCR of IFN-γ signaling-associated components in OECM-1-WT/CAL-27-WT and OECM-1-Ctx R /CAL-27-Ctx R cells. n = 3 (each with two technical replicates). Data are presented as mean ± SD. Statistical significance was determined using unpaired Student’s t test. ∗ p < 0.05; ∗∗∗ p < 0.001; ns, not significant. (C) Representative western blot analysis of IFN-γ signaling-related proteins in OECM-1-WT/OECM-1-Ctx R and CAL-27-WT/CAL-27-Ctx R cells. GAPDH was the loading control. The experiments were performed in triplicate. (D) Heatmap showing STAT1 and STAT3 protein levels from mass spectrometry in OECM-1 cells after cetuximab treatment (500 μg/mL) across different passages. (E) Representative western blot analysis of STAT family in OECM-1-WT/OECM-1-Ctx R and CAL-27-WT/CAL-27-Ctx R cells. α-tubulin was used as the loading control. The experiments were performed in triplicate. (F) Representative western blot analysis of STAT1 protein levels in OECM-1 cells across different passages of cetuximab treatment (500 μg/mL). GAPDH was used as a loading control. The experiments were performed in triplicate. (G) Left: Schematic of the mouse experiment. Murine oral squamous cell carcinoma MOC-L2-1 cells were transduced with a doxycycline (DOX)-inducible vector for the knockdown of Stat1 (shStat1) or a scramble control (shScr) and were then inoculated subcutaneously into C57BL/6 mice. Doxycycline administration was initiated on day 18 to induce vector expression in syngeneic tumors. Mice were treated with either isotype IgG or murine anti-PD1 (200 μg) for 8 doses at specified time points. Right: Tumor growth curves are presented as mean ± SD. n = 3 per group. Statistical significance was determined using unpaired Student’s t test. ∗∗ p < 0.01. (H) Upper: Histogram showing weights of shScr and shStat1 MOC-L2-1 tumors. n = 3 per group. Statistical significance was determined using unpaired Student’s t test. ∗ p < 0.05. Lower: Representative images of tumors. See also .
Techniques Used: Quantitative RT-PCR, Gene Expression, Western Blot, Control, Mass Spectrometry, Transduction, Plasmid Preparation, Knockdown, Expressing
![Tyrosine 701 phosphorylation promotes STAT1 degradation in cetuximab-resistant HNSCC (A) Upper: Representative western blot analysis of STAT1 protein levels in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells following treatment with cycloheximide (20 μg/mL) for the indicated times. β-actin was the loading control. Lower: Quantification of STAT1 protein levels. Data are presented as the mean ± SD. n = 3 per group. Statistical significance was determined using unpaired Student’s t test. ∗ p < 0.05; ∗∗∗ p < 0.001; ns, not significant. (B) Upper: Representative western blot analysis of STAT1 protein levels in OECM-1-Ctx R (left) and CAL-27-Ctx R (right) cells transfected with STAT1 (OECM-1-Ctx R -STAT1 and CAL-27-Ctx R -STAT1) and treated with proteasome inhibitor (MG132, 20 μM) for 18 h. Snail was the positive control for proteasomal degradation. Lower: Representative western blot analysis of STAT1 protein levels in OECM-1-Ctx R (left) and CAL-27-Ctx R (right) cells transfected with STAT1 (OECM-1-Ctx R -STAT1 and CAL-27-Ctx R -STAT1) and treated with lysosomal inhibitor (bafilomycin A1, 100 nM) or autophagic degradation inhibitor (hydroxychloroquine [HCQ], 20 μM). LC3B is a marker for monitoring autophagy. GAPDH was the loading control. The experiments were performed in triplicate. (C) Representative immunoprecipitation and western blot analyses of polyubiquitinated STAT1 in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells transfected with STAT1. The cells were treated with MG132 (20 μM) for 6 h to inhibit proteasome degradation. The experiments were performed in triplicate. (D) Representative western blot analysis of total STAT1, Tyr701-phosphorylated STAT1, and Ser727-phosphorylated STAT1 in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells transfected with STAT1. The cells were treated with MG132 (10 μM) for 16 h to inhibit proteasome degradation. GAPDH was the loading control. The experiments were performed in triplicate. (E) Representative immunoprecipitation and western blot analyses of polyubiquitinated STAT1 in OECM-1-Ctx R cells transfected with wild-type (WT) or Tyr701-unphosphorylatable mutant (Y701F) STAT1. Cells were treated with MG132 (10 μM) for 6 h to inhibit proteasomal degradation. The experiments were performed in triplicate. See also .](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1663/pmc12711663/pmc12711663__gr4.jpg "Tyrosine 701 phosphorylation promotes STAT1 degradation in cetuximab-resistant HNSCC (A) Upper: Representative western ...")
Figure Legend Snippet: Tyrosine 701 phosphorylation promotes STAT1 degradation in cetuximab-resistant HNSCC (A) Upper: Representative western blot analysis of STAT1 protein levels in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells following treatment with cycloheximide (20 μg/mL) for the indicated times. β-actin was the loading control. Lower: Quantification of STAT1 protein levels. Data are presented as the mean ± SD. n = 3 per group. Statistical significance was determined using unpaired Student’s t test. ∗ p < 0.05; ∗∗∗ p < 0.001; ns, not significant. (B) Upper: Representative western blot analysis of STAT1 protein levels in OECM-1-Ctx R (left) and CAL-27-Ctx R (right) cells transfected with STAT1 (OECM-1-Ctx R -STAT1 and CAL-27-Ctx R -STAT1) and treated with proteasome inhibitor (MG132, 20 μM) for 18 h. Snail was the positive control for proteasomal degradation. Lower: Representative western blot analysis of STAT1 protein levels in OECM-1-Ctx R (left) and CAL-27-Ctx R (right) cells transfected with STAT1 (OECM-1-Ctx R -STAT1 and CAL-27-Ctx R -STAT1) and treated with lysosomal inhibitor (bafilomycin A1, 100 nM) or autophagic degradation inhibitor (hydroxychloroquine [HCQ], 20 μM). LC3B is a marker for monitoring autophagy. GAPDH was the loading control. The experiments were performed in triplicate. (C) Representative immunoprecipitation and western blot analyses of polyubiquitinated STAT1 in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells transfected with STAT1. The cells were treated with MG132 (20 μM) for 6 h to inhibit proteasome degradation. The experiments were performed in triplicate. (D) Representative western blot analysis of total STAT1, Tyr701-phosphorylated STAT1, and Ser727-phosphorylated STAT1 in OECM-1-WT/OECM-1-Ctx R (left) and CAL-27-WT/CAL-27-Ctx R (right) cells transfected with STAT1. The cells were treated with MG132 (10 μM) for 16 h to inhibit proteasome degradation. GAPDH was the loading control. The experiments were performed in triplicate. (E) Representative immunoprecipitation and western blot analyses of polyubiquitinated STAT1 in OECM-1-Ctx R cells transfected with wild-type (WT) or Tyr701-unphosphorylatable mutant (Y701F) STAT1. Cells were treated with MG132 (10 μM) for 6 h to inhibit proteasomal degradation. The experiments were performed in triplicate. See also .
Techniques Used: Phospho-proteomics, Western Blot, Control, Transfection, Positive Control, Marker, Immunoprecipitation, Mutagenesis
Figure Legend Snippet: Reduced transcriptional activity of STAT1 in cetuximab-resistant HNSCC via Lys637 acetylation (A) Representative western blot analysis of the indicated proteins in OECM-1-WT/OECM-1-CtxR (left) and CAL-27-WT/CAL-27-Ctx R (right) cells transfected with STAT1 and treated with or without IFN-γ (100 ng/mL) for 24 h. α-tubulin was the loading control. The experiments were performed in triplicate. (B) Mass spectrometric analysis of CAL-27-Ctx R cells, identifying acetylation at Lys637 of STAT1. (C) Sequence alignment showing the conservation of STAT1 Lys637 across various species. (D) Representative western blot analysis of CAL-27-Ctx R and OECM-1-Ctx R cells transfected with wild-type or unacetylatable mutant STAT1(K637R), treated with or without IFN-γ (100 ng/mL) for 24 h. GAPDH was the loading control. The experiments were performed in triplicate. (E) Representative co-immunoprecipitation and western blot analyses detecting lysine-acetylated STAT1 in CAL-27-Ctx R and OECM-1-Ctx R cells transfected with wild-type STAT1 or STAT1(K637R). The cells were treated with MG132 (10 μM) for 16 h. The experiments were performed in triplicate. (F) Representative electrophoretic mobility shift assay assesses the DNA binding of wild-type STAT1 or STAT1(K637R) in CAL-27-Ctx R cells. The cells were transfected with the corresponding vectors, treated with MG132 (10 μM, 16 h) and IFN-γ (100 ng/mL, 30 min). (G) Representative western blot analysis of the indicated proteins in U3A cells transfected with STAT1(K637R) or STAT1(K637Q) mutants and treated with IFN-γ (100 ng/mL) for 24 h. GAPDH was a loading control. The experiments were performed in triplicate. (H) Representative blot detecting dimerized STAT1 and Tyr701-phosphorylated STAT1 in U3A cells transfected with STAT1(K637R) or STAT1(K637Q) mutants treated with IFN-γ (100 ng/mL) with or without disuccinimidyl suberate (DSS) (2.5 μM) for 10 min. The experiments were performed in triplicate. See also .
Techniques Used: Activity Assay, Western Blot, Transfection, Control, Sequencing, Mutagenesis, Immunoprecipitation, Electrophoretic Mobility Shift Assay, Binding Assay
Figure Legend Snippet: IFN-β and TNF-α as potential upstream regulators of STAT1 inactivation in cetuximab-resistant HNSCC (A) Schematic representation of the identification of upstream regulators using Ingenuity Pathway Analysis in OECM-1-Ctx R and CAL-27-Ctx R cells (left). Expression levels of the indicated genes based on RNA sequencing in OECM-1-Ctx R and CAL-27-Ctx R cells compared to parental cells (right). (B) ELISA of IFN-β (left) and TNF-α (right) concentrations in conditioned media from CAL-27 and CAL-27-Ctx R cells ( n = 3, with two technical replicates each). Data are presented as mean ± SD. Statistical analyses were performed using unpaired Student’s t test. ∗∗∗ p < 0.001. (C) Representative western blot of the indicated proteins in CAL-27-Ctx R cells transfected with STAT1 (CAL-27-Ctx R -STAT1) and treated with MG132 (10 μM) combined with JAK1 (left), JAK2 (middle), and TYK2 inhibitors (right) at the indicated concentrations for 16 h. GAPDH was a loading control. The experiments were performed in triplicate. (D) Representative western blot of the indicated proteins in CAL-27-Ctx R (left) and OECM-1-Ctx R (right) cells transfected with STAT1 (CAL-27-Ctx R -STAT1 and OECM-1-Ctx R -STAT1) and treated with MG132 (10 μM) and IFN-β-neutralizing antibody at indicated concentrations for 16 h. GAPDH was the loading control. The experiments were performed in triplicate. (E) Representative western blot of STAT1 Tyr701 phosphorylation in OECM-1-Ctx R (left) and CAL-27-Ctx R (right) cells transfected with STAT1 (CAL-27-Ctx R -STAT1 and OECM-1-Ctx R -STAT1) and treated with MG132 (10 μM) combined with an IFN-α-neutralizing antibody at indicated concentrations for 16 h. α-tubulin was the loading control. Experiments were duplicated. (F) Representative co-immunoprecipitation and western blot analyses to investigate the interaction between STAT1 and histone acetyltransferases in the CAL-27-Ctx R and OECM-1-Ctx R cells transfected with STAT1 (CAL-27-Ctx R -STAT1 and OECM-1-Ctx R -STAT1). The cells were then treated with MG132 (10 μM) for 16 h. The experiments were performed in triplicate. (G) Representative in vitro acetylation assay. Biotin-labeled synthetic peptides, corresponding to the sequence encompassing STAT1 lysine 637 (K637) or a mutant variant where K637 was substituted with arginine (K637R), were utilized. These peptides were incubated in the presence or absence of the histone acetyltransferase (PCAF) and with acetyl-coenzyme A (acetyl-CoA). Following the incubation, the reaction products were analyzed by dot blot for assessing acetylation levels. The experiments were performed in triplicate. See also .
Techniques Used: Expressing, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Control, Phospho-proteomics, Immunoprecipitation, In Vitro, Acetylation Assay, Labeling, Sequencing, Mutagenesis, Variant Assay, Incubation, Dot Blot
![( A ) Representative immunoblots and densitometric quantification of <t>Stat1</t> Ser727 phosphorylation levels in MLECs isolated from Ifr1 fl/fl control mice pre- and post-LPS. ( B ) Representative immunoblots and densitometric quantification of Stat1 Ser727 phosphorylation levels in MLECs in Ifr1 fl/fl control mice and endothelial cell-specific Irf1 knockout mice at baseline. ( C ) Representative immunoblots and densitometric quantification of Stat1 Ser727 phosphorylation and Irf1 protein levels in the indicated MLEC cell lines following LPS (10 μg/ml) treatment. ( D ) Representative immunoblots and densitometric quantification of STAT1 Ser727 phosphorylation and IRF1 protein levels in the indicated HLMVEC cell lines following LPS (10 μg/ml) treatment. ( E ) Schematic of the human IRF1 promoter region depicting the two highly-conserved STAT1 binding site at −44 ∼ −56 bp and −177 ∼ −188 bp. The WT and MUT GAS1 sequences used in panel ( G ) are provided. ( F ) HLMVECs subjected to vehicle or LPS conditions for 8 hours, followed by ChIP-qPCR assays for detection of STAT1 binding to the two binding sites within the IRF1 promoter region. (G) HLMVECs co-transfected with control or STAT1 plasmid along with one of three luciferase (Luc) reporter gene constructs. Schematic representations of Luc constructs are indicated. All experiments: n = 6 mice or 6 independent biological replicates per group. Data represented as means ± SDs. * P <0.05, ** P <0.01 [(A, C, D) one-way ANOVA with Bonferroni post-hoc tests, and Log-rank Mantel-Cox tests; (B) two-tailed Student’s t -tests; (E, H, I) two-way ANOVA with Bonferroni post-hoc tests].](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1169/pmc10011169/pmc10011169__cs-137-cs20220876-g5.jpg)
