phycoerythrin pe mouse anti human cxcr4 fusin monoclonal antibody  (R&D Systems)

Journal: British journal of haematology

Article Title: The BTK Inhibitor Ibrutinib (PCI-32765) Blocks Hairy Cell Leukaemia Survival, Proliferation and BCR Signalling: A New Therapeutic Approach

doi: 10.1111/bjh.12867

Figure Lengend Snippet: (A) Left: overlay histogram plots that depict the mean CXCR4 intensity florescence ratio (MIFR) in the HCL cell lines, ESKOL and HC-1, and CD19-gated primary cells, as indicated above the histograms. Light grey shaded area represents the corresponding isotype control and dark grey line indicates CXCR4 expression. Right: MIFR for CXCR4 for 13 primary HCL cases; the mean and SEM are indicated by the bars and error lines respectively. (B) Representive immunoblots from HCL primary cells (HCL14, HCL15 and HCL16) stimulated for 10 min with CXCL12 in the presence or absence of 0.1 μM, 0.5 μM or 1 μM ibrutinib as indicated. pERK refers to the phosphorylated form of the protein ERK. GAPDH was used as a protein loading control.

Article Snippet: Flow cytometric analysis was done on HCL cell suspensions (ESKOL, HC-1 and primary HCL cells: HCL1 to HCL7 and HCL9 to HCL14) stained with phycoerythrin (PE)-conjugated mouse anti-human CD184 (CXCR4), allophycocyanin (APC)-conjugated mouse anti-human CD19, and isotype control PE-conjugated mouse IgG2a,κ (BD Pharmingen).

Techniques: Expressing, Western Blot

Journal: British journal of haematology

Article Title: The BTK Inhibitor Ibrutinib (PCI-32765) Blocks Hairy Cell Leukaemia Survival, Proliferation and BCR Signalling: A New Therapeutic Approach

doi: 10.1111/bjh.12867

Figure Lengend Snippet: Upon antigen stimulation, BCR signalling induces LYN and SYK phosphorylation (P in orange circles refer to phosphorylation) that initiate a signalling cascade causing downstream BTK activation. BTK is recruited into the signalling complex at the plasma membrane via the docking of its pleckstrin homology domain to phosphatidylinositol 3,4,5 triphosphate (PIP3). BTK becomes phosphorylated and therefore induces calcium release, which initiates a signalling cascade with the consequent induction of cell proliferation and survival. BTK is also involved in signalling of chemokine receptor CXCR4, which is important in cell migration and could be a suitable target for ibrutinib.

Article Snippet: Flow cytometric analysis was done on HCL cell suspensions (ESKOL, HC-1 and primary HCL cells: HCL1 to HCL7 and HCL9 to HCL14) stained with phycoerythrin (PE)-conjugated mouse anti-human CD184 (CXCR4), allophycocyanin (APC)-conjugated mouse anti-human CD19, and isotype control PE-conjugated mouse IgG2a,κ (BD Pharmingen).

Techniques: Activation Assay, Migration