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. 2015 May 7;6(5):e1743. doi: 10.1038/cddis.2015.96

Figure 2.

Figure 2

(a–e) Response of mitotic-spindle checkpoint-proficient human lung cancer cell lines, A549 and NCI-H1299, to BPT. Flow cytometric analysis of asynchronous cells show that majority of cells are arrested in the G2/M phase of cell cycle (4n DNA content) in both the cell lines. A549 untreated (a), treatment with IC50 concentration of BPT for 24 h (b) and treatment with IC70 concentration of BPT for 24 h (c); NCI-H1299 untreated or control (d) and treatment with IC50 concentration of BPT for 24 h (e). (f–m) Analysis of NCI-H358 cells using flow cytometer. Cells in the G2/M and G1/S phase synchronized by nocodazole and hydroxyurea, respectively, were released either in the fresh medium or in the fresh medium containing IC50 concentration of BPT, which exhibit greater tendency to block the cell growth at the G2/M phase. For nocodazole block experiment, figure show untreated or control cells (f), treated with 1 μM nocodazole for 18 h (g), treated with 1 μM nocodazole for 18 h and released in the fresh medium (h) and treated with 1 μM nocodazole for 18 h and released in the presence of BPT, IC50 (i). For hydroxyurea block experiment, the figure shows untreated or control cells (j), treated with 250 μM hydroxyurea for 18 h (k), treated with 250 μM hydroxyurea for 18 h and released in the fresh medium (l) and treated with 250 μM hydroxyurea for 18 h and released in the presence of BPT, IC50 (m). (n–q) Selective apoptosis in SV40-transformed cells by BPT was analysed by FACS. BNL CL2 (mouse embryonic normal hepatic cells) when exposed to BPT for 48 h at IC50 and IC70 concentrations show prominent G2/M arrest. As seen in the figure, untreated cells (n), treated with IC50 concentration of BPT, 48 h (o). BNL SV A.8 (mouse embryonic SV40-transformed hepatic cells) underwent apoptotic cell death upon incubation with BPT. The apoptosis was quantitated by measuring the % of cells that appeared in sub-G1 peak; 43 and 51% cells were found in sub-G1 peak after 48 h exposure with BPT, IC50 (q) concentration. The untreated cells (p) do not show any apoptosis