(A) FCM assay for apoptosis: 4T1 and MDA-MB-231 cells were treated in either the absence or presence of lobaplatin (10 µg/ml) or an indicated temperature for 1 h. (B) 4T1 and MDA-MB-231 cells were treated with 0, 10, 20 µg/ml lobaplatin for 24 h. (C) 4T1 and MDA-MB 231 cells were treated with microwave hyperthermia at 37, 39, 41 and 43°C for 1 h, respectively. (D) 4T1 and MDA-MB 231 cells were treated in either the absence or presence of lobaplatin (10 µg/ml) or at an indicated temperature. (E,H) Relative expression of cleaved PARP and caspase 3 expression of 4T1 and MDA-MB-231 cells were treated with 0, 10, 20 µg/ml lobaplatin for 24 h. (F,I), Relative expression of cleaved PARP and caspase3 expression of 4T1 and MDA-MB 231 cells were treated with microwave hyperthermia at 37, 39, 41 and 43°C for 1 h. (G,J) Relative expression of cleaved PARP and caspase 3 expression of 4T1 and MDA-MB 231 cells were treated in either the absence or presence of lobaplatin (10 µg/ml) or at an indicated temperature. Twenty-four hours later, PARP Caspase 3, LC3B and P62 were detected on Western blot analysis. β-actin was used to confirm that the proteins equal in each lane. Means and standard deviations in the bar charts were from three independent experiments.

ns means not significant, *P<0.05, **P<0.01, based on Student’s test. Data are presented as the mean ± SD.