Fig. 4
ZFP161 mediates the RPA-ATRIP interaction. a, b Chromatin fractionations from HCT116 cells in presence or absence of ZFP161 following 10 mM HU treatment for 2 h were used for immunoblotting with indicated antibodies. c Whole cell lysates from HCT116 cells were used for immunoprecipitation with ZFP161 antibody. d The schema of ZFP161 constructs used to identify the minimal region required for its interaction with RPA32. e HEK293T cells were transfected with S-tagged RPA32 and ZFP161 vectors. After 48 h, the interaction between ZFP161 and RPA32 was determined by immunoprecipitation and immunoblotting with indicated antibodies. f, g Whole cell lysates from HCT116 cells were used for immunoprecipitation with ZFP161 (f), or ATRIP (g) antibodies. h The schema of ZFP161 constructs used to identify the minimal region required for its interaction with ATRIP. i HEK293T cells were transfected with the indicated ZFP161 vectors. Aftter 48 h, ZFP161 and ATRIP interaction was determined by immunoprecipitation and immunoblotting with indicated antibodies. j, k HCT116 cells expressing or lacking ZFP161 were transfected with constructs encoding Myc-ATRIP, ZFP161 (WT) and D3 mutant (j) or T2 mutant (k). Whole cell lysates were immunoprecipitated with Myc antibody and immunoblotted with indicated antibodies. l ZFP161 deficient HCT116 cells were incubated with EdU and HU. Replication fork proteins were isolated by iPOND and immunoblotted with indicated antibodies. m Biotin labeled single fork DNA was incubated with HCT116 whole cell lysates and pulled down with streptavidin beads. The interaction of ATRIP and RPA on DNA was determined by immunoblotting. < : ZPF161 full length and mutants bands. Source data are provided as a Source Data file.
