Figure 5
From: Toxicity of oxidized phospholipids in cultured macrophages

Apoptotic effects of oxPL in RAW264.7 cells and BMM. Cells were incubated with the indicated concentrations of oxPLs in media under low serum conditions (0,1% FCS) for 4 h and analyzed by flow cytometry (see materials and methods). The fraction of apoptotic cells was determined from Annexin V staining of externalized phosphatidylserine. Control cells were incubated with 1% v/v Ethanol (EtOH). 2,5 mM H2O2 or 1 μM staurosporin (STS) in the incubation media, which were used as reference agents inducing necrosis or apoptosis, respectively. Panel A: 50 μM POVPC and PGPC induce apoptosis in RAW264.7 macrophages. PGPC is a more potent inducer of cell death than POVPC under these conditions. Results are means of 6 replicates out of three independent experiments. Panel B: 50 μM PGPC and POVPC induce apoptosis in BMM, which are slightly more sensitive to the oxPL compared with RAW264.7 macrophages. Results are means from one representative experiment of 4 replicates of one pooled sample (BMM isolated and pooled from femurs and tibias of 5 mice, see materials and methods). Panel C: 50 μM 1-O-alkyl ether analogs of PGPC and POVPC (E-PGPC and E-POVPC, respectively) induce apoptosis in RAW264.7 macrophages. Panel D: Representative histogram of RAW264.7 cells after exposure to 1% v/v Ethanol (EtOH), 1 μM staurosporin (STS), POVPC or PGPC. Experimental conditions as described under Panel A. All Results are expressed as means +/− SD. Significance was determined by Student’s t-test (two tailed, unpaired). ** p ≤ 0,01, *** p ≤ 0,005.