Macrophages play a major role in atherosclerotic plaque destabilization, leading to acute coronary syndromes and sudden death. Removal of macrophages from plaques might therefore represent a promising approach to stabilize plaques. Recently, we showed that the protein synthesis inhibitor cycloheximide selectively depleted macrophages from rabbit atherosclerotic plaques. However, another protein synthesis inhibitor, puromycin, induced apoptosis of both macrophages and smooth muscle cells (SMCs). The mechanism of action of these two translation inhibitors is different and could account for the dissimilar effects on SMC viability. The question remains whether selective depletion of macrophages is a specific characteristic of cycloheximide or that other translation inhibitors, with a mechanism of action close to cycloheximide, can also selectively deplete macrophages from atherosclerotic plaques. We therefore investigated the effects of anisomycin on macrophages and SMCs. Anisomycin induced apoptosis of macrophages and to a lesser extent of SMCs in vitro as well as in vivo in rabbit atherosclerotic plaques. The Jun N-terminal Kinase (JNK) phosphorylation inhibitor SP600125 had no effect on anisomycin-induced macrophage or SMC death. The p38 MAPK inhibitor SB202190 decreased anisomycin-induced p38 phosphorylation, increased extracellular signal-regulated kinases (ERK) 1/2 phosphorylation and had no effect on JNK phosphorylation in both cell types. SB202190 partially prevented anisomycin-induced macrophage, but not SMC apoptosis. Taken together, anisomycin selectively depletes macrophages in rabbit atherosclerotic plaques by apoptotic cell death. p38 MAPK and/or ERK1/2 play a major role in anisomycin-induced macrophage, but not SMC apoptosis. Therefore, anisomycin-induced apoptosis might be differentially regulated in macrophages and SMCs.