Apoptotic cell-mediated signaling events are dominant over those mediated by necrotic cells mediated signaling in BM macrophages
Serum-starved BM macrophages were pre-exposed to mixtures of apoptotic (Apo) and necrotic (Nec) BUMPT cells for 30 min. A, the number of necrotic cells was kept constant at a necrotic cell to macrophage ratio of 1:1, and the number of apoptotic cells was increased as shown. B, the number of apoptotic cells was kept constant at an apoptotic cell to macrophage ratio of 1:1, and the number of necrotic cells was increased as shown. Nonadherent cells were removed by rinsing, and macrophage lysates were probed with anti-active ERK1/2, p38, JNK, and p90Rsk antibodies. Equal loading was confirmed by Ponceau S staining of blotted proteins as well as probing for total ERK1/2, p38, JNK, or p90Rsk (supplemental Fig. S4, except for total JNK, which is not shown). Apoptotic and necrotic cells alone contributed no active ERK1/2, p38, JNK, or p90Rsk in these experiments.