Figure 6

Bortezomib modulates miR-29b expression levels in MM cells. (a) Annexin V-staining of NCI-H929 cells 24 h after treatment with vehicle or bortezomib (indicated as bort) 2, 5, or 10 nM. ^•P<0.05. (b) Immunoblot of Sp1 in NCI-H929 cells treated for 24 h with vehicle or bortezomib (indicated as bort) 5, 10, or 20 nM. GAPDH was used as loading control. Quantitative RT–PCR of miR-29b in (c) NCI-H929 cells treated with vehicle or 2, 5, or 10 nM bortezomib for 24 h or (d) SKMM1and U266 treated with 10 nM bortezomib or vehicle for 24 h. Raw Ct values were normalized to RNU44 housekeeping snoRNA and expressed as ΔΔCt. *P<0.01; ^•P<0.05. (e) Quantitative RT–PCR of miR-29b in CD138⁺ cells immunopurified from peripheral blood of a PCL patient, 24 h after systemic treatment with 1.3 mg/kg bortezomib. Normalization was performed using RNU44 snoRNA and ΔΔCt calculations. *P<0.01. (f) Quantitative RT–PCR of miR-29b in NCI-H929 treated with vehicle (DMSO) or 5 nM bortezomib (indicated as bort) or with 5 nM bortezomib and 20 μM Z-IETD-FMK for 24 h. *P<0.01; ^•P<0.05