ESW attenuates cardiomyocyte apoptosis by upregulation of phosphorylation of Akt and survivin followed by upregulation of Bcl2 in an in vivo mouse model. (a) Immunofluorescence staining to detect Bcl2 and survivin in various regions of the mice myocardium. After fixation, the tissues were stained with anti-Bcl2 antibody (yellow) and anti-survivin antibody (red), while nuclei were stained with DAPI (blue). The levels of Bcl2 and survivin expression were compared among the three groups (control, YM155, DOX, ESW + DOX, and YM155 + ESW + DOX) using confocal microscopy. Representative images are shown (magnification 400×; scale bars, 20 μm). (b) The hearts were harvested at 14 days after the DOX injection and examined by Western blot for p-Akt, Akt, survivin, Bcl2, and GAPDH. The expression levels were normalized to GAPDH (internal control) and are indicated relative to those of the control (n = 8/group). Representative images from at least three independent results are shown. *Significant difference compared to control (p < 0.05). †Significant difference compared to the DOX condition (p < 0.05). ‡Significant difference compared to the ESW + DOX group (p < 0.05). The English in this document has been checked by at least two professional editors, both native speakers of English. For a certificate, please see: http://www.textcheck.com/certificate/ycCcrH.