Fig. 7.
In-cell DNA fragmentation effect of HLBT-100 against the following cell lines: A375, A459 and MDA MB231 was evaluated. 1–2 × 105 cells were seeded on glass coverslip in 12 well cell culture plate (costar, USA) and treated with drug. An equal vol of 10% formalin was added in each well for 10–15 min at RT. The plate was centrifuged at 3000g for 20 min to spin down all the cells. Liquid was discarded and cells were treated with 0.1% Triton X100 for 5 min and removed from triton. Cells were treated with DAPI (0.5 μg/ml) in 0.1% BSA for 5 min and washed 3× with normal saline. Finally, the cells were mounted with mowiol (sigma) and transferred onto microscopic glass slides, dried in dark and visualized under fluorescence microscope (Nikon and Zeiss, USA). Effect of HLBT-100 on fragmented is indicated by arrows