PCI-32765 significantly altered gene expression in MM cells and OC lineage cells stimulated with M-CSF/RANKL. (A) MM (ANBL6, INA-6) were treated with PCI-32765 (1μM) for an hour, and cell lysates were subjected to immunoblotting using specific Abs to determine the effects of the drug on Btk signaling cascade. (B) Patient MM cells (MM1, MM2) and MM1R cells were treated with PCI-32765. (C) INA6 cells, with or without IL-6, were treated with PCI-32765 overnight followed by real-time quantitative RT-PCR for MIP-1α/CCL3, which was normalized to HPRT as an internal control. Fold changes triggered by the drug were further normalized relative to the medium control. (D) Human donor OCPs stimulated with M-CSF/RANKL were treated with PCI-32765 for 7 days, and mRNA changes of these target genes were assayed by real-time quantitative RT-PCR using specific primers.