Microscopy from Scientific Research

Complemented DIAPH1-P1 fibroblasts were treated with 10 M EdU for 30 min, exposed to 100 nM CPT for 1 h and then permeabilised/fixed. EdU positive cells were labelled with Alexa-fluor-488 using click chemistry. Cells were then subjected to a PLA reaction using antibodies to DIAPH1 and H2AX. (Top) Representative images of the PLA reaction (red spots) in EdU positive (green nuclei) and negative (blue) nuclei stained with DAPI. (Bottom) PLA spots were quantified in at least 50 EdU-positive and 50 EdU-negative cells per experiment, per cell line, per condition. The mean number of PLA spots per cells from n = 3 independent experiments is shown.

DOI