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Blood, Vol. 113, Issue 13, 3040-3049, March 26, 2009
The proteasome load versus capacity balance determines apoptotic sensitivity of multiple myeloma cells to proteasome inhibition
Blood Bianchi et al.
113: 3040
Supplemental materials for: Bianchi et al
Files in this Data Supplement:
- Figure S1. MM lines with differential apoptotic sensitivity to PI (JPG, 72.8 KB)
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The MM lines U266, RPMI8226, KMS.18 and MM.1S were subjected to 48 hr treatment with increasing doses of bortezomib (Btz), and toxicity tested either by FACS apoptosis assay (upon labeling with FITC-conjugated Annexin V and propidium iodide) or by the MTT viability assay. (A) Differential PI sensitivity of the MM lines. Apoptosis is indicated as the proportion of Annexin V+ propidium iodide- cells, normalized by untreated samples. The assay was performed in triplicates at least 4 times by 4 independent investigators. One representative experiment is shown. (B) Differential decreases in viability in response to Btz in the 4 MM lines. Colorimetric determination of viability is expressed as a percentage of untreated cells. One representative experiment out of 3 is shown.
- Figure S2. Effect of patient derived stromas on proteasome activity of MMC (JPG, 56.6 KB)
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Stromal cells were purified from bone marrow aspirates as described in the Methods section. MM lines expressing differential proteasome activity were cultured onto layers of stromal cells for 2 to 4 days in a 1:5 stroma to MM ratio, purified, lysed, and assayed for proteasome specific peptidase activity (see Methods). In the experimental timeframe, proteasome activity of MM lines was not affected by exposure to stromas. One representative experiment is shown.
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