Fig. 5: GSK3β favors ROS generation in response to DMNQ.
From: GSK3β is a key regulator of the ROS-dependent necrotic death induced by the quinone DMNQ
a Cytofluorimetric analysis measuring cell death percentages (PI positivity) in the indicated U87MG cell lines, treated or not with the indicated combinations of molecules. DMNQ was used 30 µM. N-acetylcysteine (NAC) was 5 mM. Incubation was for 24 h. Data are presented as mean ± SD. n = 3. b Levels of reactive oxygen species as measured by Carboxy-H2DCFDA fluorescence in U87MG GSK3β+/+ and GSK3β−/− cells treated with 30 µM DMNQ. c Levels of reactive oxygen species as measured by ROS Deep Red Dye fluorescence in the indicated U87MG cell lines treated with 30 µM DMNQ. d Levels of reactive oxygen species as measured by ROS Deep Red Dye fluorescence in the indicated U87MG cell lines treated with 10 µM G5. e Time-course analysis of NQO1, NQO2, HMOX1 and GCLM mRNA expression levels. U87MG GSK3β+/+ and GSK3β−/− cells were treated with 30 μM of DMNQ for the indicated times and the mRNA levels were monitored by qRT-PCR. Data are from three experiments; +SD. f Immunoblot analysis of NRF2 levels in U87MG GSK3β+/+ and GSK3β−/− cells treated with 30 µM DMNQ for the indicated times. Actin was used as loading control. g Cytofluorimetric analysis measuring cell death percentages (PI positivity) in the indicated U87MG cell lines, treated or not with the indicated combinations of molecules. DMNQ was used 30 µM, Dicoumarol 100 µM, Tacrine 20 µM. Incubation was for 24 h. Data are presented as mean ± SD. n = 3.
