MMSET alters cell growth, cell cycle, and apoptosis in multiple myeloma cells. (A) MMSET affects growth of multiple myeloma cells. Cloned KMS11 cells harboring inducible shRNA were grown in the presence of doxycycline, and cells were counted at the indicated time points. Fresh media with or without doxycycline was added at days 7, 10, and 13 to allow continued cell growth. Two different clones are shown in the graphs. Knockdown using the C-terminal shRNA shows a drastic change in growth behavior. (B) MMSET alters adhesion of multiple myeloma cells. Knockdown cells where grown in the presence of doxycycline for 7 days, and pictures were imaged as described in “Adhesion assay.” (C) MMSET depletion induces apoptosis. Knockdown cells were grown in the presence of doxycycline, and annexin V staining was performed as described in “Cell cycle analysis and apoptosis assay.” (D) MMSET induces cell cycle arrest. Cell-cycle profiles of knockdown cells grown in the presence of doxycycline for 7 days were assessed after a 30-minute BrdU pulse as described in “Cell cycle analysis and apoptosis assay.” MMSET knockdown decreases the percentage of cells in S-phase, and this effect is more pronounced using the C-terminal shRNA. (E) MMSET induces proliferation. Knockout cells infected with vector control, wild-type, and mutants of MMSET were grown, and cell counts were performed over a period of 2 weeks. Cells containing wild-type and active mutant (F1177A) of MMSET proliferate faster than cells infected with vector control or the enzymatically dead MMSET (Y1118A).