Fig. 2

Effect of PDTC treatment on Nrf2 target genes, GCL activity, and glutathione content. a HO-1 and b GCLM mRNA expression levels measured by quantitative RT-PCR from primary adult astrocyte cultures and hippocampal neuron cultures after 24 h PDTC treatment. The mRNA expression levels are presented as mean relative expression normalized to β-actin ± SD. c Representative immunoblots with antibodies against Keap1, HO-1, and GAPDH from neonatal astrocytes treated with PDTC for 4 h. d Quantification of Keap1 and e HO-1 protein levels. Data are expressed as fold change over control cells normalized to GAPDH ± SD. f Neonatal astrocytes were treated with PDTC for 24 h, before GCL activity was determined. Data are expressed as mean nanomole γGC produced per minute per milligram protein ± SD. g The cellular glutathione content was measured after 24 h PDTC treatment of neonatal astrocytes and normalized to cellular protein content ± SD. Data are expressed as nanomole glutathione content per milligram protein. h Glutathione released into the media from neonatal astrocytes was measured 24 h after PDTC treatment. Data are shown as nanomole of glutathione released into the well ± SD. *p < 0.05; **p < 0.001 relative to corresponding control