Dlx-2 shRNA prevents metabolic stress-induced necrosis in MTS. (A, B) MCF-7 cells stably transfected with control and Dlx-2 shRNA were seeded into 1.2% agarose-coated 96-well plates at a density of 400 cells per well and cultured for 7, 8, and 9 days. Then the cells were dissociated and stained with HO/PI (A), and apoptotic and necrotic cells were scored. The values are expressed as mean ± SE (n = 3). *P < 0.05, **P < 0.01 versus control; #P < 0.05; ##P < 0.01 versus control shRNA (B). (C) Formation, growth, and morphology of MTSs made using MCF-7 control and Dlx-2 shRNA stable cells. To calculate MTS size, diameters of 5 spheroids were measured every day. Results are expressed as mean ± SE (n = 3). ##P < 0.01 versus control shRNA.