Figure 5
From: A Xenopus Dbf4 homolog is required for Cdc7 chromatin binding and DNA replication
XCdc7 chromatin interaction requires XDbf4. (A) Sperm nuclei were incubated for 30 min in Xenopus interphase extract previously depleted with antibodies specific to XDbd4 (XDbf4-) or with non-immune serum (NI-), minus or plus added recombinant GST-XDbf4 protein, minus or plus geminin. Chromatin was then isolated and immunoblotted with antibodies specific for XCdc7, XMcm7, XOrc1 and XCdc6. (B) Sperm nuclei were incubated for 30 min in XCdc7-depleted extracts (XCdc7-) or non-immune depleted extracts (NI-) containing recombinant GST-XCdc7, recombinant XDbf4 previously treated with thrombin to remove the GST tag (XDbf4), recombinant geminin and/or microcystin. Incubation of recombinant proteins in non-immune depleted extract lacking sperm are shown as negative controls. Chromatin was then isolated and immunoblotted for XDbf4, XCdc7, XMcm7 and XOrc1.