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. 2009 Apr 28;4(4):e5351. doi: 10.1371/journal.pone.0005351

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Sanbe et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Cytochrome c was released from mitochondria (Mito) into the cytosolic fraction (Cyto) in R120G TG mice while GGA treatment (GGA 2%) and overexpression of HSPB8 inhibited its release. (B) Immunostaining for activated caspase-3 in sections derived from hearts at 30 weeks of age. Cardiomyocytes were identified by phalloidin staining for actin (green). Activated caspase-3 (arrowheads) is apparent in the HSPB5 R120G (R120G TG) section. GGA treatment (GGA 2%) and overexpression of HSPB8 (tTA/HSPB8/HSPB5 R120G triple TG; Triple TG) reduced caspase-3 activation. (C) TUNEL assays in hearts from HSPB5 R120G TG mice at 30 weeks of age. Cardiomyocytes were identified by phalloidin staining for actin (green). Positive signals were apparent only in the HSPB5 R120G-derived sections and are indicated by an arrow. GGA treatment (GGA 2%) and overexpression of HSPB8 (Triple TG) decreased the TUNEL-positive signals. (D) Quantification of TUNEL signals. Between 5×10⁵ nuclei were counted for each sample and the percentage of TUNEL-positive cells were determined (n = 4 mice).