Research Paper Volume 8, Issue 4 pp 810—830
Mitotic degradation of yeast Fkh1 by the Anaphase Promoting Complex is required for normal longevity, genomic stability and stress resistance
Figure 8. Fkh1 and Apc5 physically associate throughout the cell cycle. (A) Fkh1-TAP cells expressing plasmid-borne GST-APC5 were grown overnight to early log phase. Lysates were prepared and IPed using antibodies against the TAP epitope. Westerns using antibodies against TAP and GST were done. (B) Fkh1-TAP cells expressing the empty GST vector were used as described for (A). (C) The various Fkh1-TAP cells expressing plasmid-borne GST-APC5 were arrested in S phase using HU. HU was then washed away, with cells released synchronously back into the cell cycle. Samples removed every 30 minutes were subjected to CoIP using TAP and Clb2 antibodies. Westerns were performed on input and bound samples using antibodies against TAP and GST. GAPDH was used as a load control. See also