CaMKII activation participates in doxorubicin cardiotoxicity and is attenuated by moderate GRP78 overexpression
Fig 4
CaMKII activation contributes to Doxorubicin (Dox) cardiotoxicity.
Neonatal rat cardiomoycytes were treated with 5μM AIP or vehicle starting 14h after the addition of 1μM Dox or medium (untreated). (A) Levels of cleaved caspase3 after 48h Dox treatment (n = 3–5). (B) Percentage of TUNEL positive cells in cells treated for 72h with Dox or medium in combination with AIP or vehicle. The ratio of DAPI positive and TUNEL positive nuclei was built. (n = 6). * P < 0.05 for matched 2-way ANOVA with Bonferroni post hoc testing. (C) Representative images of TUNEL positive (red) and negative cells. DAPI (blue) was used to stain nuclei. Scale bar represents 100 μm. (D) Number of cells positive for cleaved caspase9 after 48h treatment with Dox or ctrl in combination with AIP or vehicle (n = 5) (E) Representative images for caspase9 downregulation by AIP treatment. Scale bar represents 100 μm. (F) Apoptosis inducing factor (AIF) translocation: Relative signal intensity of anti-AIF antibody in the nucleus vs. cytosol after 48h Dox treatment (with and without AIP inhibition). All images except (B): p < 0.05, **p < 0.01 and ***p < 0.001 all for 1-way ANOVA with Bonferroni post hoc testing.