phosphoplus jak2 tyr1007 tyr1008 antibody duet  (Cell Signaling Technology Inc)

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JKA2/STAT3 pathway protein detection in HK-2 cells transfected with LncRNA NKILA overexpression virus (A) Representative western blotting bands and (B) semi-quantification of results of protein expression of phosphorylated JAK2 and STAT3 (n=3). (C) RT-qPCR statistical results (n=3). ## P<0.01 and # P<0.05 compared with the normal group. ns, not significant; p-, phosphorylated; OE, overexpression; NC, negative control; JAK, Janus kinase; RT-qPCR, reverse transcription quantitative PCR.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Over Expression, Virus, Western Blot, Expressing, Quantitative RT-PCR, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 signaling pathway-associated proteins in HK-2 cells transfected with LncRNA NKILA knockdown lentivirus (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Statistical analysis results of reverse transcription-quantitative PCR for channel indicators. ## P<0.01 compared with the normal group (n=3), **P<0.01 and *P<0.05 compared with the control + KD-NC group. JAK, Janus kinase; KD, knockdown; NC, negative control; p-, phosphorylated.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Knockdown, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 pathway protein detection in the recovery experiment of HK-2 cells treated with AG490. (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Reverse transcription-quantitative PCR for channel indicators (n=3). ## P<0.01 compared with the normal group, **P<0.01 and *P<0.05 compared with control + DMSO group and △△ P<0.01 compared with OE-NKILA group. OE, overexpression; p-, phosphorylated; JAK, Janus kinase; ns, not significant.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Over Expression

Journal: Frontiers in Pharmacology

Article Title: Baicalin-geniposide attenuates pulmonary inflammation and vascular injury via HMGB1 blockade: insights from a cerebral ischemia-reperfusion model and implications for pulmonary hypertension

doi: 10.3389/fphar.2026.1822890

Figure Lengend Snippet: Effect of BG on JAK2/STAT3/HMGB1 signaling pathway in LPS-induced BV2. (A) Quantification of the p-JAK2/JAK2 in LPS-induced BV2 (n = 3). (B) Quantification of the p-STAT3/STAT3 in LPS-induced BV2 (n = 3). (C) Quantification of the HMGB1 in LPS-induced BV2 (n = 3). (D) Quantification of the HMGB1 in the cytoplasm in LPS-induced BV2 (n = 3). (E) Quantification of the HMGB1 in the nucleus in LPS-induced BV2 (n = 3). Compared with the control group, ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05, ## p < 0.01, ### p < 0.001; compared with the BG-10 μg/mL group, △ p < 0.05, △△ p < 0.01, △△△ p < 0.001.

Article Snippet: Primary antibodies added: anti-JAK2 antibody (CellSignalingTechnology, 1:1,000), anti-STAT3 (Servicebio, 1:500), anti-P-JAK2 (CellSignalingTechnology, 1:1,000), anti-P-STAT3(Servicebio, 1:500), anti-HMGB1 (Servicebio, 1:1,000),anti-β-actin (Servicebio, 1:500),anti-β-tubulin (Proteintech, 1:10,000),anti-LaminB1 (immunoway, 1:1,000).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Baicalin-geniposide attenuates pulmonary inflammation and vascular injury via HMGB1 blockade: insights from a cerebral ischemia-reperfusion model and implications for pulmonary hypertension

doi: 10.3389/fphar.2026.1822890

Figure Lengend Snippet: Effect of BG on JAK2/STAT3/HMGB1 signalling pathway in brain tissue of MCAO rats. (A–C) Expression of HMGB1 in brain, lung tissues and serum (n = 6). (D) Quantification of the p-JAK2/JAK2 in brains (n = 3). (E) Quantification of the p-STAT3/STAT3 in brains (n = 3). (F) Quantification of the HMGB1 in brains (n = 3). (G) Quantification of the HMGB1 in the cytoplasm in brains (n = 3). (H) Quantification of the HMGB1 in the nucleus in brains (n = 3). (I) Immunofluorescence assay of HMGB1 in brain tissues. Compared with the sham group, ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05, ## p < 0.01, ### p < 0.001; compared with the BG-25 mg/kg group, △ p < 0.05, △△ p < 0.01.

Article Snippet: Primary antibodies added: anti-JAK2 antibody (CellSignalingTechnology, 1:1,000), anti-STAT3 (Servicebio, 1:500), anti-P-JAK2 (CellSignalingTechnology, 1:1,000), anti-P-STAT3(Servicebio, 1:500), anti-HMGB1 (Servicebio, 1:1,000),anti-β-actin (Servicebio, 1:500),anti-β-tubulin (Proteintech, 1:10,000),anti-LaminB1 (immunoway, 1:1,000).

Techniques: Expressing, Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Baicalin-geniposide attenuates pulmonary inflammation and vascular injury via HMGB1 blockade: insights from a cerebral ischemia-reperfusion model and implications for pulmonary hypertension

doi: 10.3389/fphar.2026.1822890

Figure Lengend Snippet: In LPS-induced BV2, BG reduces JAK2 activation and blocks the JAK2-STAT3-HMGB1 pathway, thereby reducing the inflammatory response. (A) Quantification of the p-JAK2/JAK2 in BV2 (n = 3). (B) Quantification of the p-STAT3/STAT3 in BV2 (n = 3). (C) Quantification of the HMGB1 in BV2 (n = 3). (D) Immunofluorescence assay of HMGB1. Compared with the control group, ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05; compared with the BG-50 μg/mL group, △ p < 0.05, △△ p < 0.01; compared with the CA1 group, ○ p < 0.05, ○○ p < 0.01.

Article Snippet: Primary antibodies added: anti-JAK2 antibody (CellSignalingTechnology, 1:1,000), anti-STAT3 (Servicebio, 1:500), anti-P-JAK2 (CellSignalingTechnology, 1:1,000), anti-P-STAT3(Servicebio, 1:500), anti-HMGB1 (Servicebio, 1:1,000),anti-β-actin (Servicebio, 1:500),anti-β-tubulin (Proteintech, 1:10,000),anti-LaminB1 (immunoway, 1:1,000).

Techniques: Activation Assay, Immunofluorescence, Control

Journal: Immunity, Inflammation and Disease

Article Title: Ginkgolide B Alleviates Airway Inflammation in Hyperoxia Lung Injury

doi: 10.1002/iid3.70364

Figure Lengend Snippet: Western blot detection of Jak2, Stat3, and phosphorylation. (A) Western blot images of Jak2, Stat3, p‐Jak2, and p‐Stat3 in rat lung tissue in three groups. (B) Protein relative expressions of Jak2, Stat3, p‐Jak2, and p‐Stat3 in rat lung tissue in three groups using Image J software. Six rats per group, repeated three times in each experiment, and the data were pooled. Room air ( n = 3–7), hyperoxia ( n = 6–7), hyperoxia +GB ( n = 3–6).

Article Snippet: PhosphoPlus Jak2 (Tyr1007/Tyr1008) Antibody Duet , Cell Signaling Technology TM , 8224S , 1:1000.

Techniques: Western Blot, Phospho-proteomics, Software

Journal: Journal of Hepatocellular Carcinoma

Article Title: Pleione bulbocodioides Polysaccharides Reprogram Tumor-Associated Macrophages via JAK2/STAT3 Inhibition to Suppress Hepatocellular Carcinoma

doi: 10.2147/JHC.S578276

Figure Lengend Snippet: Effects of PBPs on modulating macrophage polarization and the expression of genes related to the JAK2/STAT3 pathway in tumors. ( A and B ) M2 markers (Arg-1, and CD206); ( C and D ) M1 markers (CD86, and iNOS); ( E and F ) JAK2 and STAT3 transcripts. Data are presented as mean ± SD (n = 3).

Article Snippet: Primary antibodies against JAK2 (cat. no. AF6022), phospho-JAK2 (cat. no. AF3024), STAT3 (cat. no. AF6294), phospho-STAT3 (cat. no. AF3293), BCL-2 (cat. no. AF6139), and β-Actin (cat. no. AF7018) (Affinity Biosciences, Cincinnati, OH, USA).

Techniques: Expressing

Journal: Journal of Hepatocellular Carcinoma

Article Title: Pleione bulbocodioides Polysaccharides Reprogram Tumor-Associated Macrophages via JAK2/STAT3 Inhibition to Suppress Hepatocellular Carcinoma

doi: 10.2147/JHC.S578276

Figure Lengend Snippet: Effects of PBPs on regulating protein expression related to JAK2/STAT3 signaling, apoptosis, and angiogenesis in tumors. ( A ) Representative Western blot images; ( B ) p-JAK2/JAK2; ( C ) p-STAT3/STAT3; ( D ) BCL-2; ( E ) BAX; ( F ) VEGF. Data are presented as mean ± SD (n = 3).

Article Snippet: Primary antibodies against JAK2 (cat. no. AF6022), phospho-JAK2 (cat. no. AF3024), STAT3 (cat. no. AF6294), phospho-STAT3 (cat. no. AF3293), BCL-2 (cat. no. AF6139), and β-Actin (cat. no. AF7018) (Affinity Biosciences, Cincinnati, OH, USA).

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: Age-dependent JAK2 expression in midbrain dopaminergic neurons (A-B) Representative immunofluorescence images showing JAK2 expression in the substantia nigra pars compacta (SNc) of mice at 2, 6, 12, and 18 months of age. Sections were co-stained with antibodies against JAK2 and either tyrosine hydroxylase (TH) (A) or Nurr1 (B). JAK2 immunoreactivity was barely detectable in 2-month-old mice but became evident from 6 months of age and increased further in 12- and 18-month-old mice. Scale bar: 50 μm. (C) Quantification of relative JAK2 fluorescence intensity in the SNc at each age after normalization to background fluorescence. (D-E) Quantification of the percentage of JAK2-positive cells among TH-positive neurons (D) and Nurr1-positive neurons (E) across age groups. Data are presented as mean ± SEM.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: JAK2 V617F -induced Nurr1 transcriptional activation (A-B) Luciferase reporter assays showing the effect of JAK2 V617F , a constitutively active mutant, on the transcriptional activity of full-length Nurr1 (pCMV-fNurr1) (A) and GAL4-Nurr1(LBD) fusion protein (B) in SK-N-BE(2)C cells. (C) JAK2 V617F -induced Nurr1 activation was significantly suppressed by pharmacological JAK2 inhibitors, AZD1480 and ruxolitinib. *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA followed by Tukey’s post hoc test. (D) Co-expression of transcriptional coactivators (SRC1, SRC3, and PGC1α) further potentiated JAK2 V617F -induced Nurr1 transcriptional activity in SK-N-BE(2)C cells. ***p < 0.001; Student’s t -test. (E) JAK2 V617F -induced Nurr1 activation was not affected by Nurr1 agonists, amodiaquine (AQ) or chloroquine (CQ). (F) JAK2 V617F did not alter the transcriptional activity of RXRα, the heterodimeric partner of Nurr1.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Activation Assay, Luciferase, Mutagenesis, Activity Assay, Expressing

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: Effect of JAK V617F on transcriptional activity of other nuclear receptors (A) Luciferase reporter assay showing that JAK2 V617F significantly enhanced Nor1 transcriptional activity, whereas no effect was observed on Nur77. ***p < 0.001, Student’s t test (B-C) JAK2 V617F did not alter ligand-induced transcriptional activity of PPARα (B) or PPARγ (C) in the presence of 15d-PGJ2. (D-E) JAK2 V617F had no effect on ligand-induced transcriptional activity of the glucocorticoid receptor (GR) activated by cortisol (D) or liver X receptor (LXR) activated by 27-hydroxycholesterol (E).

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Activity Assay, Luciferase, Reporter Assay

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: Phosphorylation-independent interaction of JAK2 with Nurr1-LBD (A) Luciferase reporter assays showing that single tyrosine (Y)-to-phenylalanine (F) substitutions within the Nurr1-LBD did not abolish JAK2 V617F -induced transcriptional activation. (B) Double and triple Y-to-F mutations at putative JAK2 recognition sites predicted by a group-based phosphorylation prediction system also retained responsiveness to JAK2 V617F . (C) Immunoblot analysis using a pan–phospho-tyrosine antibody revealed no significant difference in Nurr1 tyrosine phosphorylation levels in the presence or absence of JAK2 V617F . (D) Co-immunoprecipitation analysis showing detection of JAK2 in Nurr1-LBD–associated protein complexes, indicating a physical interaction between JAK2 and Nurr1-LBD.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Phospho-proteomics, Luciferase, Activation Assay, Western Blot, Immunoprecipitation

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: JAK2-mediated stabilization of Nurr1 protein (A) Quantitative real-time PCR analysis showing relative Nurr1 mRNA levels following overexpression of mock vector, wild-type (WT) JAK2, or JAK2V617F. No significant differences in Nurr1 mRNA levels were observed among groups. (B) Immunoblot analysis and quantification of Nurr1 protein levels following overexpression of mock vector, WT JAK2, or JAK2 V617F . JAK2 V617F significantly increased Nurr1 protein levels. ***p < 0.001, Student’s t -test (C) Representative fluorescence images and quantification of nuclear Nurr1-GFP intensity in SK-N-BE(2)C cells. JAK2 V617F significantly increased nuclear Nurr1-GFP fluorescence. ***p < 0.001; one-way ANOVA with Tukey’s post hoc test (D) Cycloheximide chase assay showing that JAK2 V617F delayed Nurr1 protein degradation compared with mock-transfected cells. *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t -test

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Real-time Polymerase Chain Reaction, Over Expression, Plasmid Preparation, Western Blot, Fluorescence, Transfection

Journal: bioRxiv

Article Title: Janus kinase 2 regulates Nurr1 protein stability in dopaminergic neurons of the aging midbrain

doi: 10.64898/2026.03.19.712974

Figure Lengend Snippet: Protective effects of JAK2V617F and Nurr1 against oxidative stress–induced cytotoxicity and ROS accumulation (A) Hydrogen peroxide (H₂O₂) induced cytotoxicity in a concentration-dependent manner in SK-N-BE(2)C cells. Co-overexpression of JAK2 V617F and Nurr1 significantly attenuated H₂O₂-induced cytotoxicity. *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Tukey’s post hoc test (B) Rotenone induced cytotoxicity in a concentration-dependent manner in SK-N-BE(2)C cells. Co-overexpression of JAK2 V617F and Nurr1 significantly attenuated rotenone-induced cytotoxicity. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA, Tukey’s post hoc test (C) Representative fluorescence images and quantification of intracellular reactive oxygen species (ROS) levels measured using 2′,7′-dichlorofluorescein diacetate (DCFH-DA) following exposure to H₂O₂ or rotenone in mock-, JAK2 V617F -, Nurr1-, or JAK2 V617F + Nurr1–overexpressing cells. (D-E) Quantification of DCF fluorescence intensity showing that overexpression of JAK2 V617F or Nurr1 reduced H₂O₂-induced (D) and rotenone-induced (E) ROS accumulation. Co-expression of JAK2 V617F and Nurr1 produced an additional protective effect. *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against JAK2 (Cell Signaling Technology, Danvers, MA, USA), c-Myc (Roche Diagnostics GmbH, Mannheim, Germany), or β-actin (Abcam, Cambridge, UK).

Techniques: Concentration Assay, Over Expression, Fluorescence, Expressing, Produced