Figure 4

Dlx-2 plays a role(s) in metabolic stress-induced necrosis. (A-E) A549 cells were stably transfected with Dlx-2 shRNA. The cells were analyzed by real-time PCR for Dlx-2 expression (A), and treated with GD 12 h and analyzed using Western blotting (B). The cells were treated with GD for 18 h (C) and stained with HO/PI (D), and apoptotic and necrotic cells were scored (E). (F-J) HepG2 cells were stably transfected with Dlx-2 shRNA. The cells were analyzed by real-time PCR for Dlx-2 expression (F), and treated with GD for 12 h and analyzed using Western blotting (G). The cells were treated with GD for 18 h (H), stained with HO/PI (I), and apoptotic and necrotic cells were scored (J). (K-Q) MDA-MB-231 cells were stably transfected with 2 different Dlx-2 shRNA (T1 and T2). The cells were analyzed by real-time PCR for Dlx-2 expression (K), and treated with GD for 12 h and analyzed using Western blotting (L). The cells were treated with GD for 18 h (M), stained with HO/PI (N), and apoptotic and necrotic cells were scored (O). The cells were treated with GD for 12 h and analyzed for HMGB1 (P) and LDH release (Q). The values obtained from real-time PCR, HO/PI staining, and LDH release assay are expressed as mean ± SE (n = 3). *P < 0.05, **P < 0.01 versus control; ^#P < 0.05, ^##P < 0.01 versus control shRNA. Arrow in panels B, G, and L, a putative modified form of Dlx-2.