Fig. 4

Cl-amidine combined with docetaxel synergistically induces cell apoptosis, cell cycle arrest, and autophagy in MCF7/TamR cells. a Flow cytometric analysis of 0.1 μM docetaxel and 25 μM cl-amidine combincation accelerated apoptosis of the MCF7/TamR cells compared to either individual treatment. b Western blot analysis of the activation of caspase 3, Bcl-2, Bak, and Bad in MCF7/TamR cells treated with 0.1 μM docetaxel and 25 μM cl-amidine. GAPDH was used as an internal control. c,d Flow cytometric analysis of 0.1 μM docetaxel and 25 μM cl-amidine combination induced cell cycle arrest in the G2/M phase compared to docetaxel or cl-amidine alone. The relative quantification was shown in (d). e Immunofluorescence staining for LC3B showing 0.1 μM docetaxel and 25 μM cl-amidine combination induced large punctate structures in MCF7/TamR cells (white arrow). Nuclei were stained with DAPI. Scale bar, 50 μm. f Western blot analysis of the LC3B protein greatly accumulated under the combined treatment. GAPDH served as loading control