Fig. 4

Induction of apoptosis by treatment with LL28. a-c A549, H460, and H1299 cells were treated with LL28 (0.1 and 1 μM) for 3 days. a The chromatin condensation in the nuclei was observed by fluorescence microscopy, photographed, and counted. b The expression of cleaved PARP (Cl-PARP) was determined by Western blot analysis. c The cell cycle distribution in vehicle- or LL28-treated cells was analyzed by flow cytometry. (D-H) A549 and H460 cells were treated with linsitinib (1 μM) and dasatinib (100 nM) in combination or LL28 (1 μM) for 3 days. d Cell viability was determined by the MTT assay. e Anchorage-dependent colony forming ability was determined as described in the Methods section. f The chromatin condensation in the nuclei was observed by fluorescence microscopy, photographed, and counted. g The expression of cleaved PARP (Cl-PARP) was determined by Western blot analysis. h The cell cycle distribution in each test group was analyzed by flow cytometry. The bars represent the means ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001, as determined by a two-sided Student’s t-test. Con: control; L + D: linsitinib and dasatinib in combination