Fig. 4From: Non-benzoquinone geldanamycin analogs trigger various forms of death in human breast cancer cellsDHQ3 induced necroptosis in MDA-MB-231 cells. a Cells were treated with DHQ3 for 24 h. Cell lysates were prepared and examined using western blot analysis. b The expression of necroptosis-related proteins was analyzed by western blot. c MDA-MB-231 cells were incubated with DHQ3 for 24 h and treated with primary antibodies overnight at 4 °C. The localization of the target proteins was assessed by immunofluorescence staining. Nuclei were stained with DAPI. Scale bar is 50 μm. d Cells incubated with DHQ3 (200 μM) for 24 h, with or without 1 h pre-treatment with Nec-1 or z-VAD-fmk, were analyzed by the MTT assay. *P < 0.05. e MDA-MB-231 cells treated with DMSO, Nec-1 (20 μM), DHQ3 (200 μM) alone or with Nec-1 pre-treatment, were analyzed by Annexin V-FITC/PI assayBack to article page