Figure 5.

HOXC13 binds Cdc6, ORC1 and ORC2 in vitro. (A) GST or GST-HOXC13, immobilized on agarose beads were incubated with in vitro translated [³⁵S]-labeled Cdc6 or ORC1. Proteins retained by extensively washed beads were loaded onto a 10% acrylamide-SDS gel. The input lanes contain a fraction of the radiolabeled proteins prior to binding (IN). Graphs show the amount of bound radiolabeled proteins as % of the input. (B) [³⁵S]-labeled HOXC13 was incubated with GST, GST-Cdc6 full length, GST-Cdc6 aminoterminal or GST-Cdc6 carboxyterminal and processed as described in A. (C) [³⁵S]-labeled Cdc6 was incubated with GST-HOXC13 full length, GST-HOXC13 aminoterminal, GST-HOXC13 carboxyterminal or GST, and processed as described in A. (D) Cdc6 and HOXC13 constructs used in the assay. (E) Co-immuno-precipitation experiments performed with lysates from asynchronous crosslinked T98 G cells (Input) using the indicated antibodies for immuno-precipitation and blottings. (F) Immunodetection of endogenous ORC2 after co-immuno-precipitation with exogenous GFP-tagged HOXC13 in transiently-transfected asynchronous U2OS cells (Input).