Journal: Cell Reports

Article Title: CDK8 Fine-Tunes IL-6 Transcriptional Activities by Limiting STAT3 Resident Time at the Gene Loci

doi: 10.1016/j.celrep.2020.108545

Figure Lengend Snippet:

Article Snippet: Rabbit anti-pZAP70-Y319/pSYK-Y352-AF647 (Clone 65E4) , Cell Signaling , Cat#82975S; RRID: AB_2800004.

Techniques: Purification, Recombinant, Phospho-proteomics, Software

Journal: Thrombosis and haemostasis

Article Title: Impaired Glycoprotein VI-Mediated Signaling and Platelet Functional Responses in CD45 Knockout Mice

doi: 10.1055/s-0039-1692422

Figure Lengend Snippet: (A) Murine platelets from wild-type (WT) and CD45 knockout (KO) mice were stimulated with the glycoprotein VI (GPVI) agonist collagen-related peptide (CRP) (10 μg/mL) for 10 and 30 seconds and reaction was stopped using 1/10 volume 6.6 N HClO4. Samples were centrifuged and the pellets were washed and dissolved in sample buffer. These lysates were subjected to Western blot analysis and probed with pSyk Y525/ 526, pSyk Y352, pPLCγ2 Y759, and pPLCγ2 Y1217 antibodies. The same blot was probed with total Syk and total PLCγ2 antibodies as protein loading control in each lane. (B-F) Quantitation for phosphorylation of Syk and PLCγ2, data are mean ± standard error of the mean (SEM) (n = 4), *p < 0.05. (G) Washed platelets from WT and CD45 KO mice were stimulated or not with CRP (1 mg/mL) for 60 seconds at 37°C with stirring. The reaction was stopped with 1/10 volume 6.6N HClO4. The resultant precipitates were washed and resuspended with sample buffer prior to separation via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Following transfer to nitrocellulose, the membranes were probed for PKC substrate and total pleckstrin. (H) Quantification from (A) expressed as mean ± SEM of PKC substrate phosphorylation/pleckstrin (n = 3). *p < 0.05.

Article Snippet: Antibody for phosphotyrosine SFK Y416 (catalog #6943P), pSyk Y525/526 (catalog #2711), pSyk Y352 (catalog #2701), pPLCγ2 Y759 (catalog #3874), pPLCγ2 Y1217 (catalog #3871), β-Actin 13E5 (catalog #4970S), and PKC substrate antibody (catalog #22610) were purchased from Cell Signaling Technology (Danvers, Massachusetts, United States).

Techniques: Knock-Out, Western Blot, Quantitation Assay, Polyacrylamide Gel Electrophoresis, SDS Page