Short-term Adhesion Assay of Nalm-6 Cells

Short-term adhesion assays were performed as previously described (8 (link)). Briefly, 2.10⁴ Nalm-6 cells in suspension in PBS supplemented with 0.1% BSA were stimulated with 300 nM CXCL12 for 1 min and then added to VCAM-1-coated wells (with a 1 mg/ml solution). Cells were quickly spun down and allowed to settle for another minute at 37°C. As controls, cells unstimulated with CXCL12 and uncoated wells were used. Wells were then washed twice and the number of adherent cells was determined using the ATPlite luminescence assay kit (PerkinElmer, Waltham, MA).

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Mcheik S., Van Eeckhout N., De Poorter C., Galés C., Parmentier M, & Springael J.Y. (2019). Coexpression of CCR7 and CXCR4 During B Cell Development Controls CXCR4 Responsiveness and Bone Marrow Homing. Frontiers in Immunology, 10, 2970.

Publication 2019

ATPlite luminescence assay kit

Assays Cells Cxcl12 Luminescence assay Vcam 1

Corresponding Organization : Université Libre de Bruxelles

Other organizations : Institut des Maladies Métaboliques et Cardiovasculaires, Inserm, Université Toulouse III - Paul Sabatier, Walloon Excellence in Lifesciences and Biotechnology

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Variable analysis

independent variables

CXCL12 stimulation

dependent variables

Number of adherent cells

control variables

Nalm-6 cells in suspension in PBS supplemented with 0.1% BSA
VCAM-1-coated wells
Uncoated wells

controls

Positive control: Cells stimulated with 300 nM CXCL12
Negative control: Cells unstimulated with CXCL12

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