Phorbol 12 myristate 13 acetate

To derive BMMs, bone marrow cells were isolated from the femurs and tibiae of mice, and cultured in RPMI 1640 with L-glutamine (Sigma, cat #R8758) containing 10% FBS (Seradigm), 20% macrophage colony-stimulating factor (M-CSF)-conditioned medium, and 1% primocin (Invivogen) at 37°C with 5% CO₂. Equal volume of media was added on day 4. On day 7, cells were harvested and used for infection assays. M-CSF condition media was obtained from an L-929 fibroblast cell line (ATCC). U937 cells (ATCC) were cultured in RPMI 1640 containing 10% FBS (Seradigm) and 1% Pen-Strep (ThermoFicher) at 37°C with 5% CO₂ and were differentiated with 10ng/ml Phorbol 12-myristate 13-acetate (Adipogen) for 24hrs followed by culture in PMA-free media for 48hrs prior to infections.

FPFT-2216 and pomalidomide were synthesized by Fujimoto Pharmaceutical Group. Avadomide, iberdomide, siremadlin, idasanutlin, navtemadlin, milademetan, and MG-132 were procured from MedChemExpress; Z-VRPR-FMK and phorbol 12-myristate 13-acetate (PMA) were procured from Adipogen Life Sciences. Lenalidomide and ionomycin were obtained from Cayman Chemical, BAY 11-7082 from FUJIFILM Wako Pure Chemical Corporation, and safimaltib from Selleck Chemicals. Rituxan Intravenous Infusion (rituximab) was obtained from Zenyaku Kogyo. Z-VRPR-FMK was dissolved in PBS, and other reagents were dissolved in DMSO. The final DMSO concentration used for in vitro experiments did not exceed 0.25%.