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. 2006 Mar 1;20(5):613–623. doi: 10.1101/gad.1345006

Figure 2.

Figure 2.

Specificity in the interaction of Jab1 with E2F1. (A) Bacterially expressed GST-E2F1 interacts with 35S-labeled Jab1 in vitro. Likewise, GST-Jab1 binds with 35S-labeled E2F1. (B) Nuclear extracts from REF52 cells were immunoprecipitated with anti-E2F1 antisera (KH95) and protein G beads, separated by SDS-PAGE, and immunoblotted with anti-Jab1 antisera. In the lower panel, immunoprecipitated E2F1 was detected by immunoblotting with anti-E2F1 antisera (C-20). (C) HEK293 cells were transfected with HA-E2Fs and with pcDNA3 or pcDNA3–5×-Myc-Jab1. Myc-Jab1 complexes were immunoprecipitated with anti-Myc antisera (9E10) and protein G beads, separated by SDS-PAGE, and immunoblotted with anti-HA antisera. In the lower panel, immunoprecipitated complexes were immunoblotted with anti-Jab1 antisera to assess levels of Jab1 immunoprecipitated. (D) Lysate inputs were immunoblotted for E2F protein levels. (E) Lysates were assayed for Jab1 input levels by immunoblotting.