1. Adriamycin-induced oxidative stress, activation of MAP kinases and apoptosis in isolated cardiomyocytes 
Huiquan Lou, Kuljeet Kaur, Anita K. Sharma, Pawan K. Singal 
Pathophysiology 
Volume 13, Issue 2, Pages (May 2006)
DOI: /j.pathophys
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

2. Fig. 1
(A and B) Time course of phosphorylation of ERK1/2 and p38 MAP kinases induced by adriamycin in isolated adult rat cardiomyocytes. Cardiac myocytes were treated with 10μmol/L ADR for the indicated time duration. Each phosphorylated MAP kinase was measured by Western blot analysis with its respective phospho-specific MAP antibody. The total amount of each MAP kinase protein was examined from the same stripped membrane with the kinase specific antibody: (A) ERK1/2 and (B) p38. In each figure, upper panel shows the Western blot and lower panel shows the densitometric analysis of MAP kinase activities. The values are the ratio of phosphorylated/total MAP kinase. The results were normalized for all experiments by a random setting of the densitometry at 100% for the control cardiomyocytes. Data are mean±S.E.M. from five independent experiments done in duplicate. *P<0.05 vs. control.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

3. Fig. 2
(A and B) Dose-effect study of the phosphorylation of ERK1/2 and p38 kinases induced by adriamycin in isolated rat cardiomyocytes. Cardiac myocytes were treated with indicated concentrations of ADR for 10min for ERK1/2 and 1h for p38: (A) ERK1/2 and (B) p38. Each phosphorylated MAP kinase was measured by its specific antibody. In each figure, upper panel shows the Western blot and lower panel shows densitometric analysis of MAP kinase activities. The values are the ratio of phosphorylated/total-MAP kinase. The results were normalized for all experiments by a random setting of the densitometer for the control samples as 100%. Data are average from two experiments done in duplicate.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

4. Fig. 3
(A and B) Adriamycin (ADR) activated ERK1/2 and p38 MAP kinases with and without trolox (TRO) in isolated rat cardiomyocytes. Cardiac myocytes were pretreated with 20μmol/L trolox for 1h and then incubated with 10μmol/L ADR for 10min for ERK1/2 (A) and 1h for p38 (B). Each phosphorylated MAP kinase was measured by Western blot analysis with the specific MAP kinase antibody. The total amount of MAP kinase proteins was examined from the same stripped membrane for each MAP kinase with an anti-MAP kinase antibody. Upper panel shows the Western blots and lower panel shows the densitometric analysis of MAP kinase activities. The values are the ratio of phosphorylated/total MAPKs. The results were normalized for all experiments by a random setting of the densitometer of the control samples as 100%. Data are mean±S.E.M. from three independent experiments done in duplicate. *P<0.05 vs. control; #P<0.05 vs. ADR.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

5. Fig. 4
(A and B) Detection of apoptosis in isolated cardiomyocyte by Hoechst staining (A)—upper panel: control myocytes and lower panel: adriamycin exposed myocytes. Nuclear fragmentation due to adriamycin is shown by arrow. (B) Percentage of cardiomyocyte with nuclear fragmentation from three independent experiments done in duplicate; based on Hoechst stain. Data are mean±S.E.M. *P<0.05 vs. control.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

6. Fig. 5
Time-course of changes in Bax/Bcl-xl due to adriamycin. Isolated cardiomyocytes were treated with ADR (10μmol/L) for up to 24h. (A) Protein levels of Bax and Bcl-xl were measured by Western blot analysis with their respective antibodies at different time points. Both were detected with the same stripped membrane. (B) Densitometric analysis of Bax and Bcl-xl. The values are the ratio of Bax/Bcl-xl. The results were normalized for all experiments by a random setting of the densitometer for control as 100%. Data are mean±S.E.M. from three independent experiments done in duplicate. *P<0.05 vs. control.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

7. Fig. 6
Dose-effect study of the Bax/Bcl-xl changes due to adriamycin (0.1–10μmol/L). Isolated cardiomyocytes were treated with ADR in indicated doses for 1h: (A) Western blot analysis and (B) densitometric analysis. Data are mean±S.E.M. of three experiments. *P<0.05 vs. control.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

8. Fig. 7
Effects of trolox on adriamycin-induced changes in the ratio of Bax/Bcl-xl. Isolated cardiomyocytes were pretreated with trolox (20μmol/L) for 1h, followed by combination of ADR and TRO treatment for another 1h: (A) Western blot analysis and (B) densitometric analysis. Data are mean±S.E.M. from three independent experiments done in duplicate. *P<0.05 from all other groups; #P<0.05 from the ADR group.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions

9. Fig. 8
Proposed scheme for adriamycin-induced cardiomyocyte apoptosis and its modulation by antioxidant trolox.
Pathophysiology  , DOI: ( /j.pathophys )
Copyright © 2006 Elsevier Ireland Ltd Terms and Conditions