2.6. Parental ssDNA assay.

To visualize parental ssDNA in U2OS or FBH1 KO cells in response to Wee1i inhibition, 25,000 cells per sample were plated on circle coverslips in a 12-well plate and grown overnight. Cells were pretreated with 50 μM Idu for 48 hrs (Idu was refreshed after 24 hrs). Cells were then treated with 200 nM or 400 nM adovasterib or DMSO (vehicle control) for 8 or 24 hrs before cell fixation. Cells were stained using BrdU and PCNA antibodies following the immunofluorescence protocol.