Figure 4.

3F_ax-Neu5Ac treatment has a minimal impact on stroma-mediated bortezomib resistance in MM1S^Heca452 cell line. MM1S^Heca452 cells were treated with 300 mM 3F_ax-Neu5Ac or dimethyl sulfoxide (DMSO) [vehicle control (CTRL)] for seven days. After treatment, cells were seeded onto 80% confluent layer of HS5 expressing GFP (A), patient-derived bone marrow stromal cells (BMSC) (B) and BMEC-60 stained with Tag-it Violet™ (C) or plastic. BMEC-60 were also stimulated with 10 ng/mL TNFα for 4 hours (h) to induce activation. Cells were co-cultured for 24 h and then treated with 5 nM bortezomib for a further 24 h. After incubation, cells were collected and cell death was examined by Annexin V-APC and PI staining. One-way ANOVA was used to determine statistical significance with Dunnett’s multiple comparison post-hoc testing. *P<0.05; **P<0.01; ***P<0.001; ns: non-significant.