Fig. 4: Multiple domains within the RECQ4 N-terminus interact with APC/C.

a (left) A schematic diagram of RECQ4 wild type (WT), Q757X, and indicated N-terminal fragments. (right) Fold changes in the amount of APC1 or APC5 pulled down, normalized to the bait FLAG-RECQ4 proteins relative to FLAG-RECQ4 WT based on the western blot analysis shown in (c). b Representative western blot analysis of the indicated proteins in chromatin-bound (CB) fractions prepared from RECQ4 knockdown (KD) HEK293 cells transiently overexpressing the indicated FLAG-RECQ4 constructs. APC1 and APC5 were used as loading controls. c Representative western blot analysis of APC1 and APC5 co-purified with the indicated FLAG-RECQ4 constructs using the CB fractions shown in (b). d (left) A schematic diagram of the His-RECQ4-FLAG constructs for expression and purification from E. coli. (right) Fold changes in the amount of APC1 or APC5 pulled down from the CB fraction prepared from parental HEK293 cells normalized to the recombinant RECQ4 bait protein fragments relative to the RECQ4_1-492 fragment based on the western blot analysis shown in (e). APC/C-binding-domain 1 (ABD1) and ABD2 are labeled with red lines. e Representative western blot analysis to detect APC1 and APC5 from the CB fraction prepared from parental HEK293 cells (input; top panels) and those bound to M2 agarose in the presence of the indicated recombinant His-RECQ4-FLAG proteins (lower 3 panels). Source data are provided as a Source data file.