The assessment of post-systolic shortening (PSS) by speckle tracking echocardiography allows myocardial ischemic memory imaging. Because the endocardial layer is more vulnerable to ischemia, the assessment of this layer might be useful for detecting ischemic memory. Serial echocardiographic data were acquired from nine dogs with 2 min of coronary occlusion followed by reperfusion. Regional deformation parameters were measured in the risk and normal areas. Using speckle tracking echocardiography, circumferential strain was analyzed in the endocardial, mid-wall, and epicardial layers; and radial strain was analyzed in the inner half, outer half and entire (transmural) layers. In the risk area, peak systolic and end-systolic strain in the circumferential and radial directions significantly decreased during occlusion, but recovered to the baseline levels immediately after reperfusion in all layers. However, circumferential post-systolic strain index (PSI), a parameter of PSS, significantly increased during occlusion, and the significant increases persisted until 20 min after reperfusion in the endocardial and mid-wall layers. Radial PSI tended to increase after reperfusion in the inner half and entire layers but these increases were not significant compared with baseline. In the normal area, systolic strains and PSI in the radial and circumferential directions hardly changed before and after occlusion/reperfusion in all layers. In layer-specific analysis with speckle tracking echocardiography, circumferential PSS in the endocardial and mid-wall layers may be useful for detecting ischemic memory.