Journal: PLoS ONE

Article Title: JAK2 Exon 14 Deletion in Patients with Chronic Myeloproliferative Neoplasms

doi: 10.1371/journal.pone.0012165

Figure Lengend Snippet: Top, schematic diagram of the JAK2 protein showing JAK homology domains 1 through 7 (JH1-JH7) with the JH2 pseudokinase domain highlighted in black. The corresponding exon regions of the mRNA is shown with the exons 13, 14, and 15. Because exon 14 is consists of 88 bp, its deletion leads to frameshift and early termination of translation after coding for seven new amino acids and elimination of the V617 codon of JAK2 (lower panel). The resulting truncated JAK2 protein is shown on the bottom.

Article Snippet: A patent has been filed on finding by Quest Diagnostics and testing for JAK2 various mutations is offered at Quest Diagnostics.

Techniques:

Journal: PLoS ONE

Article Title: JAK2 Exon 14 Deletion in Patients with Chronic Myeloproliferative Neoplasms

doi: 10.1371/journal.pone.0012165

Figure Lengend Snippet: Upper panels: Detection of Δexon14 is relatively easy when the transcript is present at high levels (eg, 29% of total JAK2 transcript). Detection is more difficult when the Δexon14 transcript makes up a small proportion of total JAK2 transcript (eg, 6.9%). Normal control is shown on the bottom.

Article Snippet: A patent has been filed on finding by Quest Diagnostics and testing for JAK2 various mutations is offered at Quest Diagnostics.

Techniques: Control

Journal: PLoS ONE

Article Title: JAK2 Exon 14 Deletion in Patients with Chronic Myeloproliferative Neoplasms

doi: 10.1371/journal.pone.0012165

Figure Lengend Snippet: Prevalence and Relative Level of the ΔExon14 JAK2 Transcript in Patients with Suspected or Confirmed Myeloproliferative Neoplasms (MPNs)

Article Snippet: A patent has been filed on finding by Quest Diagnostics and testing for JAK2 various mutations is offered at Quest Diagnostics.

Techniques:

Journal: PLoS ONE

Article Title: JAK2 Exon 14 Deletion in Patients with Chronic Myeloproliferative Neoplasms

doi: 10.1371/journal.pone.0012165

Figure Lengend Snippet: Lysates were prepared from the indicated human CML K562 cell line (Lane1), a patient with chronic myelogenous leukemia (lane 2), and 5 patients with chronic MPNs (Lanes 3–7). Patient 1: non-CML CMPD, JAK2 V617F positive; Patient 2: non-CML CMPD, JAK2 V617F negative; Patient 3: non-CML CMPD, JAK2 Δexon14 positive; Patient 4: non-CML CMPD, JAK2 Δexon14 positive; Patient 5: non-CML CMPD, JAK2 Δexon14 positive. Top Panel: Probing with an anti-JAK2 N-terminal clone yielded a wild-type JAK2 band at 130 kDa in the K562 and other negative control lanes, and an additional band at 75 kDa only in patients with expression of Δexon14 transcript. Bottom Panel: An anti-JAK2 clone directed against the carboxyl-terminus of JAK2 yielded only a single band at 130 kDa.

Article Snippet: A patent has been filed on finding by Quest Diagnostics and testing for JAK2 various mutations is offered at Quest Diagnostics.

Techniques: Negative Control, Expressing

Journal: Nature communications

Article Title: EphrinB2 controls vessel pruning through STAT1-JNK3 signaling

doi: 10.1038/ncomms7576

Figure Lengend Snippet: EphrinB2 associates with SHP2, JAK2 and STAT1, and modulates STAT1 activity. (a,b) HUVEC infected with eB2-shRNA or eB2-5Y express higher levels of p-STAT1 compared to controls; p-STAT3 levels are similar. Flow cytometry (a; % positive cells is noted on each quadrant; representative of 5 experiments), immunoblotting (b);. (c) Nuclear localization of p-STAT1 (red, arrowheads) in HUVEC transduced with eB2-5Y (GFP: green); little p-STAT1 (red) is detected in HUVEC transduced with eB2-WT (GFP: green); representative images. Scale bar: 20μm (d) EphB4-Fc activates endogenous p-EphrinB and reduces p-STAT1 levels; quantitation (average fluorescence intensity/cell). Results reflect the means±SD from 3 experiments; P values from two-tailed Student t -test: N.S., non significant, * P <0.05, ** P <0.01; error bars: ±SD. (e) Serum starvation time-dependently reduces p-EphrinB levels and increases p-STAT1 levels in HUVEC. (f) EphrinB2 associates with SHP2, JAK2 and STAT1 in HUVEC; cell lysates of HUVEC were immunoprecipitated with antibodies to EphrinB2 or control IgG; precipitates and cell lysates were immunoblotted as indicated. (g–i) Quantitative analysis of EphrinB2 association with SHP2, JAK2 and STAT1 in p5 hyaloid vessels from EphrinB2 WT/WT (n=5) and EphrinB2 5Y/5Y (n=7) mice detected by PEA (top panels). Negative control: reagents alone, no cell lysate. SHP2, JAK2 and STAT1 abundance in PEA input samples (bottom). (j) PLA shows that p-EphrinB associates with SHP2 in hyaloid vessels from EphrinB2 WT/WT mice but not from EphrinB2 5Y/5Y mice. Red: EphrinB2+SHP2; blue: DAPI. Dotted line limits amplified areas in lower panels. Scale bars: 500μm (top panels), 100μm (bottom panels) (k) Quantitation of p-EphrinB+SHP2 proximity co-localization in WT hyaloid vessels regions at p5 and p7. Results (mean±SD fluorescence intensity/mm 2 area) are normalized with DAPI. Error bars: ±SD.

Article Snippet: Antibodies were labeled with Proseek Probemaker (Olink Bioscience); anti-EphrinB2 rabbit monoclonal antibody (Abcam, no. 150411) was labeled with oligo DNA probe A; anti-SHP2 rabbit monoclonal antibody (Cell Signaling Technologies, no. 3397), anti-JAK2 rabbit monoclonal antibody (Cell Signaling Technologies, no. 3230) and anti-STAT1 rabbit polyclonal antibody (Cell Signaling Technologies, no. 9172) were labeled with oligo DNA probe B according to the manufacturer’s protocol; PEA was performed using Proseek Assay Development Kit (Olink Bioscience) according to the manufacturer’s protocol.

Techniques: Activity Assay, Infection, shRNA, Flow Cytometry, Western Blot, Transduction, Quantitation Assay, Fluorescence, Two Tailed Test, Immunoprecipitation, Control, Negative Control, Amplification

Journal: Nature communications

Article Title: EphrinB2 controls vessel pruning through STAT1-JNK3 signaling

doi: 10.1038/ncomms7576

Figure Lengend Snippet: JNK3 is a target of p-STAT1 regulation. (a) HUVEC-endogenous p-STAT1 induced by serum starvation or IFNγ (10 ng/ml) stimulation specifically binds to JNK3 promoter region. Chromatin IP with p-STAT1 antibodies or control IgG; precipitated DNA was measured by qPCR with specific primers for JNK3 promoters. CM: HUVEC complete medium. Results show means (±SD shown as error bars) from 5 replicates. P values from two-tailed Student t-test: * P <0.05, *** P <0.001. (b) WT EphrinB2 represses JAK2+STAT1-driven JNK3 promoter activity; Gaussia luciferase dual-reporter assay; means (±SD shown as error bars) of 5 experiments. P values from two-tailed Student t-test: *** P <0.001. The JNK3 reporter plasmid was co-transfected in HEK293T cells with expression plasmids for JAK2+STAT1; dominant-negative (DN) JAK2+STAT1; JAK2+STAT1+eB2-WT or JAK2+STAT1+eB2-5Y. (c) Schematic representation of experimental results.

Article Snippet: Antibodies were labeled with Proseek Probemaker (Olink Bioscience); anti-EphrinB2 rabbit monoclonal antibody (Abcam, no. 150411) was labeled with oligo DNA probe A; anti-SHP2 rabbit monoclonal antibody (Cell Signaling Technologies, no. 3397), anti-JAK2 rabbit monoclonal antibody (Cell Signaling Technologies, no. 3230) and anti-STAT1 rabbit polyclonal antibody (Cell Signaling Technologies, no. 9172) were labeled with oligo DNA probe B according to the manufacturer’s protocol; PEA was performed using Proseek Assay Development Kit (Olink Bioscience) according to the manufacturer’s protocol.

Techniques: Chromatin Immunoprecipitation, Control, Two Tailed Test, Activity Assay, Luciferase, Reporter Assay, Plasmid Preparation, Transfection, Expressing, Dominant Negative Mutation

Journal: Experimental Hematology & Oncology

Article Title: Development of a high resolution melting analysis assay for rapid identification of JAK2 V617F missense mutation and its validation

doi: 10.1186/s40164-019-0134-0

Figure Lengend Snippet: a , b Melting peak in wild type and mutant sample for the JAK2 V617F mutation shows different temperature (75/53 and 75/10, respectively). c The normalized fluorescence plot which synchronization samples in maximum and minimum fluorescence emission. d Normalized and temperature-shifted difference plot of the various kinds of allele burden mutation in compare of wild type (G allele)

Article Snippet: The test and control samples were analyzed using Ipsogen JAK2 Mutant Kit (Qiagen, Germany), as the reference method.

Techniques: Mutagenesis, Fluorescence

Journal: Experimental Hematology & Oncology

Article Title: Development of a high resolution melting analysis assay for rapid identification of JAK2 V617F missense mutation and its validation

doi: 10.1186/s40164-019-0134-0

Figure Lengend Snippet: Sensitivity and specificity of Ipsogen JAK2 Mutant Kit (Qiagen, Germany) confirmed by sequencing, HRM, PCR–RFLP and, ARMS PCR methods

Article Snippet: The test and control samples were analyzed using Ipsogen JAK2 Mutant Kit (Qiagen, Germany), as the reference method.

Techniques: Mutagenesis, Sequencing

Journal: Revista Brasileira de Hematologia e Hemoterapia

Article Title: Cytogenetics, JAK2 and MPL mutations in polycythemia vera, primary myelofibrosis and essential thrombocythemia

doi: 10.5581/1516-8484.20110116

Figure Lengend Snippet: Electropherogram of the BioEdit program, showing the absence of mutations in exon 12 of the JAK2 gene

Article Snippet: No patients with PV, even the two cases who did not have the JAK2 V617F mutation, exhibited mutations in exon 12 of the JAK2 gene ( ). shows an electropherogram from the BioEdit program; there are no anomalous peaks compared with the sequence pattern and thus no differences in the sample tested.

Techniques: