stat2  (Cell Signaling Technology Inc)

Journal: Immuno

Article Title: Conditional Stat2 Knockout Mice as a Platform for Modeling Human Diseases

doi: 10.3390/immuno6010007

Figure Lengend Snippet: Generation of conditional Stat2 KO mice. ( a ) Targeting strategy used to construct Stat2 fl/fl mice with exons 5 through 8 flanked with loxP sites. E represents exons with their corresponding number. Brown boxes depict exons in wild type allele and green boxes depict exons after successful integration of targeting vector. ( b ) Confirmation of loxP sites integration in the Stat2 gene by Southern blot analysis (9.8 kb DNA fragment). ( c ) Genotyping of mouse-tail DNA by PCR confirms deletion of Stat2 floxed allele after breeding with CMV-Cre mouse.

Article Snippet: Antibodies against STAT1 (Cat#10144–2-AP), STAT2 (cat#51075–2-AP), β-Actin (cat#66009–1-Ig), HRP-conjugated anti-mouse IgG (cat#SA00001–1) and anti-rabbit-IgG (cat#SA00001–2) were purchased from Proteintech, Rosemont, IL, USA.

Techniques: Construct, Plasmid Preparation, Southern Blot

Journal: Immuno

Article Title: Conditional Stat2 Knockout Mice as a Platform for Modeling Human Diseases

doi: 10.3390/immuno6010007

Figure Lengend Snippet: Impaired IFN-I signaling after conditional deletion of Stat2 by CMV-Cre—mediated recombination. ( a ) Loss of Stat2 gene expression validated in four different organs by qRT-PCR analysis. ( b ) Western blot analyses performed on several tissues confirm global Stat2 deletion ( Stat2 Δ / Δ ) by ubiquitous CMV-Cre recombinase. ( c ) Splenocytes of the indicated genotypes were left untreated or treated with IFN-β for 6 h, and ISG expression was determined by qRT-PCR. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001. Data are presented as SEM of three independent experiments. n.s; not significant.

Article Snippet: Antibodies against STAT1 (Cat#10144–2-AP), STAT2 (cat#51075–2-AP), β-Actin (cat#66009–1-Ig), HRP-conjugated anti-mouse IgG (cat#SA00001–1) and anti-rabbit-IgG (cat#SA00001–2) were purchased from Proteintech, Rosemont, IL, USA.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Expressing

Journal: Immuno

Article Title: Conditional Stat2 Knockout Mice as a Platform for Modeling Human Diseases

doi: 10.3390/immuno6010007

Figure Lengend Snippet: Stat2 Δ / Δ mice show enhanced tumor growth. ( a ) B16-F1 or ( b ) EL4 tumor cells were injected subcutaneously in WT, Stat2KO , Stat2fl/fl and Stat2 Δ/Δ . Values are shown as mean tumor volume determined over 20 days. Representative images of individual tumors are shown. *, p ≤ 0.05; **, p ≤ 0.01; and ***, p ≤ 0.001. n = 6–8 animals/group.

Article Snippet: Antibodies against STAT1 (Cat#10144–2-AP), STAT2 (cat#51075–2-AP), β-Actin (cat#66009–1-Ig), HRP-conjugated anti-mouse IgG (cat#SA00001–1) and anti-rabbit-IgG (cat#SA00001–2) were purchased from Proteintech, Rosemont, IL, USA.

Techniques: Injection

Journal: Immuno

Article Title: Conditional Stat2 Knockout Mice as a Platform for Modeling Human Diseases

doi: 10.3390/immuno6010007

Figure Lengend Snippet: Stat2 Δ / Δ fibroblasts have impaired antiviral response to type I IFNs. ( a ) Lung fibroblasts derived from the indicated genotypes were pretreated with IFN-β for 24 h. Cells were then infected with VSV-GFP (green) at an MOI of 0.01 and imaged after 24 h. Representative bright field and fluorescent images are shown. ( b ) Western blot analysis shows STAT2 expression in fibroblasts of indicated genotypes. Representative images are shown of n = 2.

Article Snippet: Antibodies against STAT1 (Cat#10144–2-AP), STAT2 (cat#51075–2-AP), β-Actin (cat#66009–1-Ig), HRP-conjugated anti-mouse IgG (cat#SA00001–1) and anti-rabbit-IgG (cat#SA00001–2) were purchased from Proteintech, Rosemont, IL, USA.

Techniques: Derivative Assay, Infection, Western Blot, Expressing

Journal: Immuno

Article Title: Conditional Stat2 Knockout Mice as a Platform for Modeling Human Diseases

doi: 10.3390/immuno6010007

Figure Lengend Snippet: Efficient targeted deletion of Stat2 in mouse tissues. Expression of STAT2 and STAT1 was analyzed in various tissues by Western blot analysis. ( a ) Bone marrow–derived cDCs generated from Stat2 fl/fl mice crossed with CD11c-Cre mice ( Stat2 Δ-cDC ). ( b ) Colons from Stat2 fl/fl mice crossed with Cdx2-Cre mice ( Stat2 Δ-CE ). ( c ) Lungs from Stat2 Δ-cDC and Stat2 Δ-CE mice were included for specificity of targeted deletion. Wild-type (WT) and Stat2KO mice served as positive and negative controls, respectively. ACTIN was used as an internal protein loading control.

Article Snippet: Antibodies against STAT1 (Cat#10144–2-AP), STAT2 (cat#51075–2-AP), β-Actin (cat#66009–1-Ig), HRP-conjugated anti-mouse IgG (cat#SA00001–1) and anti-rabbit-IgG (cat#SA00001–2) were purchased from Proteintech, Rosemont, IL, USA.

Techniques: Expressing, Western Blot, Derivative Assay, Generated, Control

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Effect of BITC on CNOT2 and pSTAT3 and their binding in SK-Hep1 and Huh7 cells. ( A ) Overexpression of CNOT2 in liver cancer patients implies poor survival rate along with original regression ratio with STAT3. ( B ) Effect of BITC on CNOT2 and pSTAT3 in SK-Hep1 and Huh7 cells. Band intensities were quantified and normalized to β-actin. ( C ) Effect of BITC on the binding between CNOT2 and STAT3 or c-Myc in SK-Hep1 cells. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: Binding Assay, Over Expression

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Depletion of STAT3 or CNOT2 enhances apoptotic effect of BITC in SK-Hep1 cells. ( A ) Effect of STAT3 or CNOT2 depletion in BITC treated SK-Hep1 cells. ( B ) Effect of STAT3 depletion on pro-PARP and pro-caspase3 in BITC treated SK-Hep1 cells. ( C ) Effect of CNOT2 depletion on pro-PARP and pro-caspase3 in BITC treated SK-Hep1 cells. Band intensities were quantified and normalized to β-actin. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques:

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Effect of BITC on Warburg effect in SK-Hep1 and Huh7 cells. ( A ) Effect of BITC on Warburg effect proteins in SK-Hep1 and Huh7 cells. ( B ) Effect of BITC on LDH production in SK-Hep1 and Huh7 cells. STAT3 or CNOT2 depletion vs untreated control. ***p < 0.001 vs untreated control. ( C ) Effect of BITC on glucose consumption in SK-Hep1 and Huh7 cells. *p < 0.05, ***p < 0.001 vs untreated control. ( D ) Effect of pyruvate treatment or overexpression of CNOT2 or c-Myc on apoptosis and glycolysis proteins in BITC treated SK-Hep1 cells. (E) Effect of siSTAT3 or/and siCNOT2 on HK2, PKM2 and LDH in SK-Hep1 cells transfected with or without c-Myc overexpression plasmid. Cells were co-transfected with control vector, siSTAT3, siCNOT2 and siCNOT2 + c-Myc OE (overexpression) plasmids as indicated in each lane. Band intensities were quantified and normalized to β-actin. All experiments were performed using biological triplicates and independently repeated three times .

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: Control, Over Expression, Transfection, Plasmid Preparation

Journal: Scientific Reports

Article Title: CNOT2 /c-Myc/STAT3 signaling is critically involved in glycolysis mediated apoptosis of benzyl isothiocyanate in hepatocellular carcinoma

doi: 10.1038/s41598-026-38416-8

Figure Lengend Snippet: Scheme on mechanism of BITC via CNOT2/c-Myc/STAT3 signaling in HCCs.

Article Snippet: Blots were probed with primary antibodies against CNOT2 (#34,214, 1:1000; Cell Signaling Technology, MA, USA), c-Myc (ab32072, 1:1000, Abcam, Cambridge, UK), JAK1 (#3332, 1:1000; Cell Signaling Technology, MA, USA), STAT3 (#12,640, 1:1000; Cell Signaling Technology, MA, USA), phospho-JAK1 (#3331, 1:1000; Cell Signaling Technology, MA, USA), phospho-STAT3 (#4441, 1:1000; Cell Signaling Technology, MA, USA), pro-PARP (#9542, 1:1000; Cell Signaling Technology, MA, USA), pro-caspase-3 (#9662, 1:1000; Cell Signaling Technology, MA, USA), HK2 (#2106, 1:1000; Cell Signaling Technology, MA, USA) , PKM2 (#4053, 1:1000; Cell Signaling Technology, MA, USA), LDH (SC-133123, 1:1000; Santa Cruz, CA, USA) and β-actin (A1978, 1:10,000; Sigma-Aldrich, MO, USA).

Techniques: