Evaluation of JAK-STAT inhibition in JAK2V617F myeloproliferative leukocytes and endothelium

Vascular activation is characterized by increased proinflammatory, prothrombotic, and proadhesive signaling. Several chronic and acute conditions, including Bcr-abl-negative myeloproliferative neoplasms (MPNs), graft-vs-host disease, and COVID-19, have been noted to have increased activation of the Janus kinase (JAK)-signal transducer and downstream activator of transcription (STAT) pathways. Two notable inhibitors of the JAK-STAT pathway are ruxolitinib (JAK1/2 inhibitor) and fedratinib (JAK2 inhibitor), which are currently used to treat MPN patients. However, in some conditions, it has been noted that JAK inhibitors can increase the risk of thromboembolic complications. We sought to define the anti-inflammatory and antithrombotic effects of JAK-STAT inhibitors in vascular endothelial cells and leukocytes. By using endothelialized microfluidics perfused with whole blood samples, we demonstrated that endothelial treatment with JAK-STAT inhibitors prevented rolling of both healthy contro..., Chemicals and reagents
Ruxolitinib (CAS 941678-49-5, Cat#11609) and fedratinib (TG101348) were purchased from Cayman Chemical, dissolved in DMSO, and stored as 5 μl stock aliquots: 20 mM ruxolitinib at âˆ’20 °C and 10 mM fedratinib at âˆ’80 °C. Recombinant human TNF-α was obtained from R&D Systems (#210-TA). Nuclear extracts were prepared by using the kit from Cayman Chemical (#10009277) following the manufacturer’s guidelines. Protein was quantified by using the Pierce BCA Protein Assay Kit (ThermoScientific Cat#23225). Human factor X (#HCX-0050) and human factor VIIa (HCVIIA-0031) were purchased from Haematologic Technologies. Chromogenic substrate for the determination of factor Xa activity (Chromogenix S-2765, Diapharma) was reconstituted per manufacturer’s instructions and stored at 4 °C. The urokinase type plasminogen activator human chromogenic activity assay kit was purchased from Abcam (Cat#ab8915) and used following the manufacturer’s protocol. HEPES buffered saline ..., # Evaluation of JAK-STAT inhibition in JAK2V617F myeloproliferative leukocytes and endothelium

Dataset DOI: [10.5061/dryad.4qrfj6qr0](https://doi.org/10.5061/dryad.4qrfj6qr0)

## Description of the data and file structure

### Files and variables

#### File: 210607_HUVEC_0.4uMRux_20010823-18.avi

**Description:** 

| Field Name | Description | Data Type | Expected Length of the Variable | Units | Variables | Comments |
| :---------- | :----------------------------- | :-------- | :------------------------------ | :---- | :---------------------- | :--------------------- |
| Date | Date recorded | date | 6 | N/A | <br /> | yy/mm/dd |
| Cell Line | Cell line | text | 5 | N/A | <br /> | 5 alpha |
| Treatment A | Drug treatment prior to sample | text | 2\_8 ...,