Fig. 4
From: Direct binding of CEP85 to STIL ensures robust PLK4 activation and efficient centriole assembly
The N-terminal domain of STIL directly interacts with a C-terminal coiled-coil domain of CEP85. a Domain overview of human STIL and CEP85. CR conserved region; cc coiled-coil; NTD N-terminal domain. Constructs that were used in this work are indicated by red and green lines. b STIL NTD and CEP85 interact. The C-terminal region of CEP85 is required for this interaction. Western blot showing a pull-down experiment with immobilized, recombinant GST or GST-human STIL NTD and lysates from tissue culture cells overexpressing the indicated 3xFLAG-tagged CEP85 constructs. c The C-terminal region of CEP85 shows a yeast-two-hybrid interaction with human STIL NTD. Yeast transformed with the indicated bait and prey constructs were plated on SC-Leu/-Trp plates (selecting for bait and prey plasmid, left) and on SC-Ura plates (selecting for Ura promoter activation, right). d Recombinant chicken STIL NTD and human CEP85 cc4 directly interact with micromolar affinity. Typical ITC of chicken STIL NTD and human CEP85 cc4 binding at 25 °C. The resulting KD, ΔH and STIL NTD/CEP85 cc4 binding stoichiometry (N) as an average from a total of seven independent measurements are indicated (±standard deviation)
