Fig. 3

PPHs protect against H₂O₂-induced apoptosis in MC3T3-E1 osteoblastic cells. a Apoptosis was detected by Hoechst dye staining and quantified based on nuclear condensation or fragmentation (right). Representative pictures are shown from three independent experiments. Images were taken at a magnification of 400×. Scale bar, 20 μm. b Cells were stained with FITC-conjugated Annexin V and PI, followed by flow cytometric analysis. c Caspase-12 activity was analyzed in cells treated for 0, 12, 24, 36 or 48 h with 400 μM H₂O₂ in the presence or absence of 100 μg/mL PPHs. d Caspase-3 activity was measured in cells that were treated for 6 h with 400 μM H₂O₂ in the presence or absence of 100 μg/mL PPHs. e Cells were treated for 0, 2, 4, or 6 h with 400 μM H₂O₂ in the presence or absence of 100 μg/mL PPHs. Immunoblotting was performed using the indicated antibodies. Representative blots are shown from three independent experiments. Bottom panel, quantitative immunoblot data (n = 4). ^* p < 0.05 versus cells treated with H₂O₂ alone. PPHs porcine placenta hydrolysates