Supplementary Figures from Targeting Myddosome Signaling in Waldenström's Macroglobulinemia with the Interleukin-1 Receptor-Associated Kinase 1/4 Inhibitor R191

Supplementary Figure 1. BCWM. 1 (upper panel) and MWCL-1 cells (lower panel) were seeded in 96-well plates (20,000 cells/well) and treated with increasing concentrations (0.0, 0.15, 0.3125, 0.625, 1.25, 2.5, 5.0 �M) of the indicated IRAK inhibitors for 72 hours. Cell viability was then assayed using the tetrazolium reagent WST-1, and the data presented are representative of at least 3 independent experiments. Supplementary Figure 2. MWCL-1 and BCWM.1 cells were treated with the indicated concentrations of R191 for 24 hours. They were then stained either with DAPI and analyzed by flow cytometry for cell cycle analysis (A), or they were stained with ToPRO-3 and Annexin V, and analyzed by flow cytometry for apoptosis (B). Supplementary Figure 3. Peripheral blood mononuclear cells (PBMC) from two donors were seeded in 96-well plates (50,000 cells/well) and treated with increasing concentrations (0.0, 0.0075, 0.015, 0.030, 0.075, 0.15, 0.3, 0.6, 1.25, 2.5 �M) of R191 for 24 hours. Treated PBMC's were evaluated for the induction of apoptosis by staining with ToPRO-3 and Annexin V, and analyzed by flow cytometry, and the data presented are representative of 3 independent experiments. Supplementary Figure 4. (A) BCWM.1 (left) and MWCL-1 cells (right) were treated with 1.25 �M R191 for 24 hours, and extracts were studies by RPPA analysis. Expression levels of selected intermediates are shown as the means {plus minus} SD for duplicate samples, and "*" indicates a p-value of