Activated T cells were retrovirally transduced with MSCV-CXCR5-GFP and PIB-PD-1-RFP. GFP+RFP+ co-transduced cells were sorted to > 90% purity. After resting in complete RPMI-1640 media for 2-3 hours, cells were re-suspended to a density of 3 × 106 cells/ml in the same media supplemented with or without 1 μg/ml recombinant CXCL13 (Peprotech) dropped onto L-Lysine-coated slides that were pre-incubated with PD-L1-hIgGFc or control human IgG. After incubation at 37°C for 30 min, cells were fixed with 1% paraformaldehyde and stained with AF568 Phalloidin (Invitrogen) to visualize F-actin. Slides were mounted with the ProlongGold Antifade reagent (Invitrogen) and examined with an Olympus FV1000 upright microscope using × 20 air immersion lens. Cells with an elongated shape (ratio of the cell length and width > 1.5) are considered as polarized.
Recombinant cxcl13
Manufactured by Thermo Fisher Scientific
Sourced in United States
Recombinant CXCL13 is a chemokine protein produced using recombinant DNA technology. It is a member of the CXC chemokine family and plays a role in the recruitment and activation of B cells.
Automatically generated - may contain errors
2 protocols using recombinant cxcl13
1
Visualizing CXCR5 and PD-1 Dynamics
2
CXCR5+ NK Cell Migration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For the transwell migration assays, 600 μl serum-free medium containing recombinant CXCL13 (Peprotech) at 50 ng/ml was placed in the lower chamber of a 24-well transwell plate with a pore size of 8 μm (Corning, New York, NY, USA). 5 × 105 NK cells isolated from PBMCs were added to 100 μl serum-free medium in the upper chamber. The cells were then incubated for 2 h at 37 °C. The proportion of CXCR5+NK cells in the lower chamber was determined using flow cytometry. Relative migration was calculated as the frequency of CXCR5+ NK cells in the lower chambers of the CXCL13 group divided by the control group.
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