Journal: Pathology and Oncology Research

Article Title: Pathological Changes and Expression of JAK-STAT Signaling Pathway Hallmark Proteins in Rat Retinas at Different Time Points After Retinal Ischemia Reperfusion Injury

doi: 10.3389/pore.2022.1610385

Figure Lengend Snippet: Immunohistochemistry staining showing protein expression and localization of JAK2, STAT3, p-JAK2, p-STAT3, Bcl-2, and Bax in control, RIRI 0, 6, 24, 72, and 144 h groups. (A) Protein expression of JAK2, STAT3, p-JAK2, p-STAT3, Bcl-2, and Bax in six groups. Scale bar = 50 μm. (B–G) Analysis of JAK2 (B) , STAT3 (C) , p-JAK2 (D) , p-STAT3 (E) , Bcl-2 (F) , and Bax (G) in six groups. N = 5 sections for measuring per group. Compared with control group; ns, no significance, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies were as follows: JAK2 (bs-23003r, Bioss), STAT3 (60199-1-lg, Proteintech), p-JAK2 (ab32101, Abcam), p-STAT3 (sc-8059, Santa), Bcl-2 (ab194583, Abcam), and Bax (A19684, ABclonal).

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Pathology and Oncology Research

Article Title: Pathological Changes and Expression of JAK-STAT Signaling Pathway Hallmark Proteins in Rat Retinas at Different Time Points After Retinal Ischemia Reperfusion Injury

doi: 10.3389/pore.2022.1610385

Figure Lengend Snippet: The hallmarker protein expression of JAK-STAT signaling pathway in control, RIRI 0, 6, 24, 72, and 144 h groups. (A) Protein expression of JAK2, p-JAK2, STAT3, and p-STAT3 in six time points groups. (B–E) Analysis of JAK2 (B) , p-JAK2 (C) , STAT3 (D) , and p-STAT3 (E) in six groups. N = 3–6 repeats per group. Compared with control group, ns, no significance, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies were as follows: JAK2 (bs-23003r, Bioss), STAT3 (60199-1-lg, Proteintech), p-JAK2 (ab32101, Abcam), p-STAT3 (sc-8059, Santa), Bcl-2 (ab194583, Abcam), and Bax (A19684, ABclonal).

Techniques: Expressing

Journal: Oncogene

Article Title: An activating mutation in the transmembrane domain of the granulocyte colony-stimulating factor receptor in patients with acute myeloid leukemia.

doi: 10.1038/sj.onc.1205767

Figure Lengend Snippet: Figure 5 Tyrosine phosphorylation of (a) anti-JAK2 and (b) anti-G-CSFR immunoprecipitates from Ba/F3 transfectants sti- mulated with or without 100 ng/ml G-CSF for 3 min or 10 ng/ ml mIL-3 for 10 min. Lysates were immunoprecipitated with anti-JAK2 or anti-G-CSFR antibodies; blots were probed with the phospho-tyrosine specific antibody PY99, stripped and re- probed with antibody against JAK2 or G-CSFR

Article Snippet: Western blots were probed with PY99, a phosphotyrosine specific antibody (Santa Cruz Biotechnology), then stripped and re-probed with antibody against JAK2 or the receptor.

Techniques: Phospho-proteomics, Immunoprecipitation

Journal: Discover oncology

Article Title: Hepatoma cell-derived exosomal SNORD52 mediates M2 macrophage polarization by activating the JAK2/STAT6 pathway.

doi: 10.1007/s12672-024-01700-y

Figure Lengend Snippet: Fig. 3 SNORD52 mediates M2 macrophage polariza- tion and activates JAK2/ STAT6 pathway. A Following transfection of OE-SNORD52/ OE-NC in THP-1 and U937 for 48 h, the mRNA expression of SNORD52 was detected by qRT-PCR. ****P < 0.0001 vs. OE-NC. B Following trans- fection of OE-SNORD52/ OE-NC in THP-1 and U937 for 48 h, the protein levels of Arginase-1 and CD163 were determined through Western blotting. *P < 0.05, **P < 0.01, ***P < 0.001 vs. OE-NC. C The ratio of CD206-positive or CD163-positive cells following transfection of OE-SNORD52/ OE-NC was analyzed by flow cytometry. ***P < 0.001 vs. OE-NC. D Following transfec- tion of OE-SNORD52/OE-NC in THP-1 and U937 for 48 h, the levels of JAK2/STAT6 pathway-related proteins were determined through Western blotting. ***P < 0.001 vs. OE-NC. E The IL-4 and IL-13 treated THP-1/U937 mac- rophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the mRNA expression of SNORD52 was detected by qRT-PCR. *P < 0.05, **P < 0.01, ***P < 0.001 vs. ASO-NC. F The IL-4 and IL-13 treated THP-1/U937 mac- rophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the protein levels of Arginase-1 and CD163 were determined through Western blotting. ***P < 0.001 vs. ASO-NC. G The ratio of CD206-positive or CD163-positive cells following transfection of SNORD52- ASO-1/-2 or ASO-NC was analyzed by flow cytometry. ***P < 0.001 vs. ASO-NC. (H) The IL-4 and IL-13 treated THP-1/ U937 macrophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the levels of JAK2/STAT6 pathway-related proteins were determined through Western blotting. ***P < 0.001 vs. ASO-NC. ns: no significance

Article Snippet: Meanwhile, the primary antibodies Arginase-1, CD163, and STAT6 used in Western blotting were also purchased from Proteintech, while other primary antibodies including JAK2, p-JAK2, p-STAT6, exosome-positive markers (CD81 and TSG101), and the corresponding HRP-conjugated secondary antibodies were available from Abcam (Cambridge, UK).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Incubation

Journal: Discover oncology

Article Title: Hepatoma cell-derived exosomal SNORD52 mediates M2 macrophage polarization by activating the JAK2/STAT6 pathway.

doi: 10.1007/s12672-024-01700-y

Figure Lengend Snippet: Fig. 5 Huh7 cells-derived exosomal SNORD52 mediates M2 macrophage polarization through activating JAK2/STAT6 pathway. A Following the establishment of co-culture model, AG-490 (a specific JAK2 inhibitor) was added and the levels of JAK2/STAT6 pathway-related pro- teins in receptor THP-1 macrophages were measured by Western blotting. B Following the establishment of co-culture model, AG-490 (a specific JAK2 inhibitor) was added and the ratio of CD206-positive or CD163-positive THP-1 macrophages was analyzed by flow cytometry. **P < 0.01, ***P < 0.001. ns: no significance

Article Snippet: Meanwhile, the primary antibodies Arginase-1, CD163, and STAT6 used in Western blotting were also purchased from Proteintech, while other primary antibodies including JAK2, p-JAK2, p-STAT6, exosome-positive markers (CD81 and TSG101), and the corresponding HRP-conjugated secondary antibodies were available from Abcam (Cambridge, UK).

Techniques: Derivative Assay, Co-Culture Assay, Western Blot, Flow Cytometry

Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation 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: 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

Journal: Cells

Article Title: The Regulatory Effects of JAK2/STAT3 on Spermatogenesis and the Redox Keap1/Nrf2 Axis in an Animal Model of Testicular Ischemia Reperfusion Injury

doi: 10.3390/cells12182292

Figure Lengend Snippet: The effect of AG490 on the phosphorylation of JAK2 and STAT3. ( A ) Representative pictures of Western blots demonstrating the protein levels of the phosphorylated forms of JAK2 and STAT3. ( B ) Diagrammatic blot demonstrating the signal intensity quantification of the phosphorylation ratios of p-JAK2/JAK2 and p-STAT3/STAT3. The values are presented as the mean ± SD ( n = 6/group), p -value < 0.05. * tIRI compared to sham and # AG490-treated compared to tIRI. Abbreviations: JAK = Janus kinase; STAT = signal transducer and activator of transcription; tIRI = testicular ischemia reperfusion injury; I = Ipsilateral.

Article Snippet: The primary antibodies for JAK2 (ab108596) and ph-JAK2 (ab195055) were purchased from abcam (Cambridge, UK).

Techniques: Western Blot