Journal: Cell Death & Disease
Article Title: HIV gp120- and methamphetamine-mediated oxidative stress induces astrocyte apoptosis via cytochrome P450 2E1
Figure Lengend Snippet: Caspase-3 activation is involved in the MA and/or gp120-mediated apoptosis in astrocytes. ( a and b ) SVGA cells were treated with MA and/or gp120 for 24 h and the cytoplasmic proteins were obtained, which were used to measure procaspase-3 (35 kDa) and cleaved caspase-3 (17 kDa) protein expressions ( a ) and caspase-3 cleavage activity ( b ). The cells were also treated with either 100 μ M vit. C ( c ), 25 μ M DAS ( d ), 50 nM DPI ( e ), or 50 nM DFO ( f ) 1 h before the treatment with MA and/or gp120 and the effect on caspase-3 cleavage activity was measured. The activity was normalized with basal levels in untreated control and reported as change in the activity. The bars represent mean±S.E. of three independent experiments with each treatment in triplicates. The P -value ≤0.05 (*) and ≤0.01(**) were considered statistically significant using two-tailed Student's t -test and multiple analysis of variance (ANOVA). The significant values shown above the bars were calculated using Student's t -test for comparison between the treatment and mock-transfected control
Article Snippet: Specific antibodies against ferritin heavy chain (B12), CYP2E1 (H-21), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (FL-335) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), antibody against caspase-3, which detects procaspase3 (35 kDa) and cleaved caspase-3 (17 kDa) was purchased from Cell Signaling Technology (Danvers, MA, USA) and antibody against CYP2D6 and CYP2B6 were purchased from Abcam (Cambridge, MA, USA).
Techniques: Activation Assay, Activity Assay, Two Tailed Test, Transfection