Fig. 5

Apoptosis analysis by TUNEL staining and cleaved caspase-3 immunofluorescence staining. Significant numbers of apoptotic cells were observed in the central regions of the tumor in Group B than Group A, while there was no significant difference in apoptosis cells between Groups C and D. A TUNEL staining, the apoptotic nuclei were stained green and the nuclei were stained blue with DAPI. B Cleaved-Caspase-3 immunofluorescence staining, when the cell was stimulated by apoptosis, the activated Cleaved-Caspase 3 in the cytoplasm was indicated as green. The nuclei were stained blue with DAPI. C Histogram of the percentage of TUNEL positive nuclei. D Histogram of the integrated density of Cleaved-Caspase 3 positive area. CS cell sheet, CE cell suspension, E ectopic tumor, O orthotopic tumor, P peripheral area, C central area. *: p < 0.05. Scale bar: 50 μm