Journal: Oncotarget

Article Title: Low-dose bortezomib increases the expression of NKG2D and DNAM-1 ligands and enhances induced NK and γδ T cell-mediated lysis in multiple myeloma

doi: 10.18632/oncotarget.13979

Figure Lengend Snippet: Annexin V and 7-AAD double negative cells were gated and analyzed for surface marker expression. ( A ) Dose response: RPMI-8226 and U266 cells were exposed to increasing doses of bortezomib for 12 h. ( B ) Time course: RPMI-8226 and U266 cells were incubated with low-dose (10 nM) bortezomib for increasing durations of time. ( C ) Representative western blot analysis of MICA, MICB, Nectin-2, and PVR expression in low-dose bortezomib-treated MM cells. β-actin was used as a loading control of cell lysates. ( D ) qRT-PCR analysis of MICA, MICB, Nectin-2, and PVR expression in RPMI-8226 and U266 cells exposed to low-dose bortezomib for 12 h. Data are expressed as relative expression of MICA, MICB, Nectin-2, and PVR mRNA in low-dose bortezomib-treated MM cells over that of normal saline-treated control cells (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: MM cells were incubated with 0, 5, 10, and 20 nM bortezomib for 12 h and MM cells were incubated with 10 nM bortezomib for 12, 24, and 36 h. Cells were then stained with Nectin-2-PE (BD Biosciences), PVR-PE, MICA-PE, and MICB-APC (R&D Systems, Minneapolis, MN, USA).

Techniques: Marker, Expressing, Incubation, Western Blot, Quantitative RT-PCR