Fig. 3: GSK3β is activated by DMNQ treatment.
From: GSK3β is a key regulator of the ROS-dependent necrotic death induced by the quinone DMNQ
a U87MG cells were treated with 30 µM DMNQ or 10 µM G5 for the indicated times. Cellular lysates were generated and immunoblots were performed with the indicated antibodies. In parallel cell death was scored by cytofluorimetric analysis and it is showed at the bottom. b U87MG GSK3β+/+ and GSK3β−/− cells were treated or not with 30 µM DMNQ for 24 h. Immunofluorescence analysis was performed to visualize mitochondria morphology, using anti-SMAC antibodies (red). Nuclei were stained with Hoechst 33258 (cyan). Bar 10 µm. Confocal images are shown in pseudocolors and were acquired with a Leica SP8 LSM. c Immunoblot analysis of SMAC levels in the indicated U87MG cells treated with 30 µM DMNQ for the indicated time points. Actin was used as loading control. d Cytofluorimetric analysis of TMRM fluorescence. The indicated U87MG cells were incubated for 30 min with TMRM (1 µM). FCCP (10 µM) was used for 5 min. Data are from three experiments. Columns represent the percentage of initial intensity of TMRM fluorescence + SD.
