Tg003

Cells were treated with 50 µM TG003 (Selleckchem, S7320) prepared in DMSO, control samples were treated with equal volumes of DMSO. Cells were treated for 24 hours prior to harvest for western blot or RNA extraction.

Proteomic/phosphoproteomic/RNA-seq experiments were performed at a cell density of 1 E+6 cells mL⁻¹. For timecourse studies, ~20 E+6 cells were grown in complete media for each timepoint (0, 8, 16, and 24 h), whereas for single-timepoint experiments, 15–20 E+6 cells in light SILAC media were treated with drug compound and cells in heavy SILAC media (L-Lysine-¹³C₆,¹⁵N₂, L-Arginine-¹³C₆,¹⁵N₄ (Cambridge Isotope, CNLM-291-H-1, CNLM-539-H-1) were treated with DMSO for 24 h. One-to-3 E+6 cells were set aside for RNA-seq. Cells were washed in phosphate-buffered saline (PBS) and cell pellets were frozen in liquid nitrogen (LN₂) and stored at −80 °C. One biological replicate for the timecourse experiment, two biological replicates for each single-timepoint condition (with a third only for RNA-seq), and three biological replicates for all AP-MS were gathered and analyzed. Single-timepoint experiments for MM.1S treated with 5 nM bortezomib or 50 μM lenalidomide or combination treatment of 15 nM Cfz and 50 μM TG003 (Selleckchem, S7320) or 15 nM Cfz and 50 μM z-VAD-FMK (Ubiquitin-Proteasome Biotechnologies, F7111) were carried out in 3 biological replicates.

G150 (cat. no. HY‐128583; MedChemExpress, NJ, USA); H‐151 (cat. no. S6652, Selleck, Houston, TX, USA); TBK1/IKKε‐IN‐2 (cat. no. S0425, Selleck); Stattic (cat. no. S7024; Selleck); diABZI (cat. no. S8796; Selleck); G140 (cat. no. HY‐133916; MedChemExpress); RU.521 (cat. no. S6841; Selleck); TG003 (cat. no. S7320; Selleck), DOX (cat. no. S1208, Selleck); etoposide (cat. no. HY‐13629, MedChemExpress).