Antibodies and reagents
Cell culture and transfection
SILAC labeling
Nuclear extraction
Protein extraction and trypsin digestion
HPLC fractionation
Kcr peptide enrichment
LC-MS/MS analysis
Database searching and protein quantification
Bioinformatics analysis
Immunoprecipitation and Western blotting
Purification of recombinant proteins
GST pull-down assay
Detection of ssDNA
Immunofluorescence staining and confocal microscopy
DNA-protein binding assay
Electrophoretic mobility shift assay
Colony formation assay
Flow cytometry
Statistical analysis
The complementary DNA (cDNA) for WT RPA1 was amplified by polymerase chain reaction and ligated into pcDNA3.1(−) plasmid containing 3×FLAG tag. RPA1 mutants including K88R, K379R, K595R, and 3KR were generated using the QuikChange Lightning Site-Directed Mutagenesis Kit. The siRNAs were purchased from the GenePharma Company of Shanghai. The target sequences were as follows: RPA1 siRNA, 5′-UUAUCAUCAAGCAGGAAUUAU-3′; CDYL siRNA, 5′-CAGAGAAUAACUCACUAAATT-3′.