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. 2022 Jul 1;14(4):925–944. doi: 10.1016/j.jcmgh.2022.06.008

Figure 2.

Figure 2

HNE induces hepatocyte death.A, The activation of caspase 3 was detected using NucView 488 Caspase-3 Substrate (green square) and cell death of HepG2 cells was visualized using propidium iodide (red square). Blue, nucleus; green, activated caspase 3, red, dead cell; yellow, merge. B, HepG2 cells viability was assayed 0, 2, 6, 12, and 24 hours after the addition of 6.25, 12.5, 25, 50, and 100 μM HNE. C, Statistical analysis of HepG2 cells viability 24 hours after the addition of 0, 6.25, 12.5, 25, 50, and 100 μM HNE is shown. D, Statistical analysis of HepG2 cells viability 2, 6, 12, and 24 hours after the addition of 100 μM HNE is shown. E, 3000, 6000, 12000, and 24000 HepG2 cells viability was assayed 24 hours after addition of 100 μM HNE. F, Cell death of HepG2 was assessed by staining cells with Annexin V and ethidium homodimer III (EthD-III), and analyzed by the flow cytometry. The analyses were done 6 hours after the addition of 25, 50, or 100 μM HNE. G, The difference in the frequency of double staining HepG2 cells with Annexin V and EthD-III 6 hours after the addition of 0, 25, 50, or 100 μM HNE is shown. H, Effect of Alda-1 on the viability of 40 μM HNE-treated HepG2 cells is shown. I, The activation of caspase 3 was detected using NucView 488 Caspase-3 Substrate (green square) and cell death of Huh-7 cells was visualized using propidium iodide (red square). Blue, nucleus; green, activated caspase 3; red, dead cell; yellow, merge. J, Huh-7 cells viability was assayed 0, 2, 6, 12, and 24 hours after the addition of 6.25, 12.5, 25, 50, and 100 μM HNE. K, Statistical analysis of Huh-7 cells viability 24 hours after the addition of 0, 6.25, 12.5, 25, 50, and 100 μM HNE is shown. L, Statistics analysis of Huh-7 cell viability 2, 6, 12, and 24 hours after the addition of 100 μM HNE is shown. M, 3000, 6000, 12000, and 24000 Huh-7 cells viability was assayed 24 hours after addition of 100 μM HNE. N, Cell death of Huh-7 was assessed by staining cells with Annexin V and EthD-III and analyzed by flow cytometry. The analyses were done 6 hours after the addition of 25, 50, or 100 μM HNE. O, The difference in the frequency of double staining Huh-7 cells with Annexin V and EthD-III 6 hours after the addition of 0, 25, 50, or 100 μM HNE is shown. P, Effect of Alda-1 on the viability of 40 μM HNE-treated Huh-7 cells is shown. C–E, H, K–M, P, The experiment was repeated 3 times, and blue, red, and green indicate the first, second, and third data, respectively.