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. 2017 Sep 28;8(55):93878–93898. doi: 10.18632/oncotarget.21317

Figure 2. Juglanin induced cytotoxic effects and apoptosis in lung cancer cell lines.

Figure 2

Up, lung cancer cell lines of (A) A549, (B) HCC827, and (C) H1975 were administered with juglanin at different concentrations, ranging from 0 μM to 80 μM for 24 h. Then the cell viability was measured via MTT analysis. And the lung normal cell (D) MRC-5 was also treated with various concentrations of juglanin as indicated for 24 h. Then, MTT assays were conducted to calculate cell viability. Down, the lung cancer cell lines of (A) A549, (B) HCC827, and (C) H1975, and the lung normal cell of MRC-5 were treated with the presented concentrations of juglanin for 48 h, followed by MTT assays. The data are presented as mean ± S.E.M. of three independent experiments performed in duplicate. * P < 0.05, ** P < 0.01 and *** P < 0.001 compared to Control group without any treatment. (E) A549 and H1975 cells were cultured with 40 μM juglanin for 24 h. Then, flow cytometry assay was conducted to assess DNA contents in the sub-G1 phase. Con: the control group without any treatments. The representative images were exhibited. The quantification of the apoptotic (F) A549 and (G) H1975 cells in sub-G1 phase was calculated through statistic analysis. A549 and H1975 cells were treated with juglanin (10, 20 and 40 μM) for 24 h. The DNA fragmentation assay was conducted to determine DNA fragmentation in (H) A549 and (I) H1975 cells. (J) The representative images of apoptotic morphology of cancer cells treated with or without 40 μM juglanin for 24 h were shown. And the number of apoptotic cells per field was quantified. (K) Transmission electron microscopy was used to observe the chromatin condensation and the apoptotic cells after 40 μM juglanin treatment for 24 h. (L) The representative image of lung cancer cells experiencing apoptosis was displayed. The quantification of (M) A549 and (N) H1975 cells in apoptosis was assessed. The data are presented as mean ± S.E.M. of three independent experiments performed in duplicate. * P < 0.05, ** P < 0.01 and *** P < 0.001 compared to Control group (Con) without any treatment.