In vitro B cell proliferation and apoptosis
To track B cell growth rate, 2 × 106 purified splenic
B cells were labeled with 5 μM Cell Trace Violet (CTV; C34557, Thermo
Fisher) before being seeded into 96-well plates with 1 ml complete
RPMI1640 medium (Gibco, C11875500BT) containing 10% FBS (Gibco,
10091-148), 10 μM HEPES (Gibco, 1804629) and 50 μM β-mercaptoethanol
(M2650, Sigma Aldrich). Next, cells were stimulated with 5 μg/ml
lipopolysaccharide (LPS) or CPG for 72 h. After
collecting cell, FCM was performed.