Journal: Febs Letters

Article Title: The N2-Src neuronal splice variant of C-Src has altered SH3 domain ligand specificity and a higher constitutive activity than N1-Src

doi: 10.1016/j.febslet.2015.05.033

Figure Lengend Snippet: N-Srcs have enhanced levels of Y416 auto-phosphorylation in vitro and in cells compared to C-Src. (A) 200 nM HisΔ80C-, N1- or N2-Src was incubated at 30 °C with 0.5 mM ATP for 0, 1 or 3 h and processed for immunoblotting with anti-pY416 and anti-His (loading control). pY416 immunoreactivity was obtained by densitometry with ImageJ and normalised to the anti-His signal. Data are plotted as mean ± S.E.M., from n = 3 independent experiments. (B) B104 cells were transfected with plasmids encoding FLAG-tag alone (control) or C-terminal FLAG-tagged full length C-, N1- or N2-Src, and the lysed after 48 h. The lysates were analysed by immunoblotting with anti-FLAG (FLAG), anti-phosphotyrosine (pY), anti-pY416 (active Src), anti-pY527 (inactive Src) and β-actin (loading control). The immunoblots are representative of at least three independent experiments.

Article Snippet: Rabbit polyclonal anti-Src pY527 and anti-Src pY416 were from Cell Signaling Technologies (Hitchin, UK).

Techniques: Phospho-proteomics, In Vitro, Incubation, Western Blot, Control, Transfection, FLAG-tag

Journal: The American Journal of Pathology

Article Title: Src Kinase Is Biphosphorylated at Y416/Y527 and Activates the CUB-Domain Containing Protein 1/Protein Kinase C δ Pathway in a Subset of Triple-Negative Breast Cancers

doi: 10.1016/j.ajpath.2019.10.017

Figure Lengend Snippet: Antibodies and Dilutions Used for IHC and Western Blot Analysis

Article Snippet: Antibody dilutions for each method are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Antibody information, manufacturer/catalog no. Western blot dilution IHC dilution Antigen retrieval ∗ FOXA1 Abcam/ab173287 NA 1:4000 1 Vimentin Agilent/M0725 NA 1:2000 2 Sox10 Bio SB/BSB2583 NA 1:100 1 CK5/6 Thermo Fisher Scientific/180267 NA 1:50 2 CDCP1 Cell Signaling Technology/4115 1:1000 1:100 1 CDCP1_pY743 Cell Signaling Technology/14965 1:1000 1:800 1 CDCP1_pY734 Cell Signaling Technology/9050 1:1000 NA NA Src Cell Signaling Technology/2108 1:1000 NA NA Lck Cell Signaling Technology/2752 1:1000 NA NA Lyn Cell Signaling Technology/2732 1:1000 NA NA SFK_pY416 Cell Signaling Technology/2101 1:1000 1:200 1 SFK_pY527 Cell Signaling Technology/2105 1:1000 NA NA PKCδ Cell Signaling Technology/2058 1:1000 NA NA PKCδ_pY311 Abcam/ab76181 1:2500 1:3000 1 N-cadherin Cell Signaling Technology/13116 NA 1:200 1 Slug Cell Signaling Technology/9585 NA 1:100 1 AMPKα Cell Signaling Technology/2532 1:1000 NA NA AMPKα_pT172 Cell Signaling Technology/2535 1:1000 NA NA β-Actin Leica Biosystems/A5441 1:5000 NA NA CD3 Novocastra/NCL-L-CD3-565 NA 1:50 2 PD1 Cell Signaling Technology/86163 NA 1:200 2 PD-L1 Cell Signaling Technology/13684 NA 1:400 2 CTLA4 Santa Cruz Biotechnology/sc-376016 NA 1:400 2 Open in a separate window AMPK, AMP-activated protein kinase; CDCP, CUB-domain containing protein; CK5/6, cytokeratin 5/6; CTLA4, cytotoxic T-lymphocyte protein 4; FOXA1, forkhead box protein A1; IHC, immunohistochemistry; Lck, tyrosine-protein kinase Lck; Lyn, tyrosine-protein kinase Lyn; NA, not applicable; PD1, programmed cell death protein 1; PD-L1, programmed cell death 1 ligand 1; PKC, protein kinase C; Sox10, Sry-box transcription factor 10.

Techniques: Western Blot