Anti bcma pe

CP-25 was supplied by the Chemistry Laboratory of Institute of Clinical Pharmacology of Anhui Medical University (Hefei, Anhui Province, China). Rituximab (Roche Pharma, Switzerland); Etanercept (Shanghai Guojian Pharmaceutical, Co., Ltd., China); BAFF (Pepro Tech, United States); anti-CD19-APC/PE/FITC, anti-CD20-FITC, anti-CD27-PE/APC, anti-BAFFR-APC, anti-BCMA-PE, and anti-TACI-PE (Biolegend, United States); anti-β-actin (ZSGB-BIO, China); Anti-TRAF2 (Santa Cruz, China); Anti-Phospho-NF-κBp65, anti-p100/52, and anti-Phospho-p38 MAPK (Cell Signaling Technology, United States); RPMI 1640 medium (Gibco, United States); fetal bovine serum (Zhejiang Tianhang Biotechnology, China).

After pre-incubation with an FcR-blocking reagent, cells were labeled for 30 min on ice with mAb anti-BAFF-R FITC, anti-BCMA PE, and anti TACI APC (Biolegend, San Diego, CA, USA). As positive control for the expression level of BLyS receptors, we used B lymphocytes labeled with mAb anti-CD19 and anti-BLyS receptor mAbs as described above. Appropriate isotype controls were used. Acquisition was performed on a FACS Calibur and included 10,000 events for apoptosis and 50,000 events for BLyS receptors. Data were analyzed using the CellQuest Pro software (BD Immunocytometry Systems; San Jose, CA, USA); the results were expressed as mean fluorescence intensity (MFI).