Seahorse analysis
Assays using the Seahorse XF 24 cell metabolism analyzer (XFe24,
Seahorse Bioscience, Billerica, MA, USA) were performed following the
manufacturer’s instructions. Purified WT and CCR2 KO B cells were
pre-stimulated with 10 μg/ml LPS (L2880, Sigma Aldrich, St. Louise, CA,
USA) for 1 h and detected in XF medium under basal conditions.
Subsequently,
1.5 μM oligomycin (abs42024304, Absin Bioscience, Shanghai, China), 1 μM
fluoro-carbonylcyanide phenylhydrazone (FCCP) (C2920, Sigma Aldrich) and
500 nM rotenone (R8875, Sigma Aldrich) plus 1 μM antimycin A
(abs42013402, Absin Bioscience) were sequentially added, at the
appropriate time of detection. Oxygen consumption rates (OCR) were
quantified to reflect the cellular energy metabolism level.