Figure 2.

Restoration of native CT positions after washing off cisplatin. Normal human dermal fibroblasts (NHDFs) were treated with 25 µM cisplatin to induce DNA damage for 4 hours. The cisplatin (Cp) was then washed off (0 h post Cp wash-off) and the cells were then allowed to repair the damage by placing them in fresh medium and samples were collected at varying time-periods post cisplatin wash-off (as indicated in Fig. 1). 2D-FISH was then performed for a set of chromosomes and at least 100 images per sample were analyzed via IMACULAT.⁵⁰ The software basically divides the nuclei of each cell in 5 shells of equal area (shell 1 being the most peripheral shell and shell 5 the innermost shell). The amount of FISH probe in each shell is then normalized with the amount of DNA and histograms are plotted. The positions of chromosomes 11 and 19 (panels A–H) and 12 and 17 (panels I–P) were determined at respective time points using 2D-FISH analyses. Cp = cisplatin. Adapted by permission from BioMed Central: reference 15, copyright 2013.