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. 2009 Sep 22;37(21):7072–7084. doi: 10.1093/nar/gkp777

Figure 7.

Figure 7.

Cdk2 is acetylated by PCAF and GCN5 in vivo. (A) Cells were transfected with Flag-cdk2 alone or together with three different acetylases (p300, PCAF or GCN5). Cell extracts were subjected to IP with anti-Flag and WBs were performed with anti-Flag and anti-Acetyl-K. (B) Cells were transfected with Flag-cdk2 or Flag-cdk2K33R alone or together with HA-GCN5. Then, cell extracts were subjected to IP with anti-Flag followed by WB with anti-Flag and anti-Acetyl-K. (C) C2C12 cell extracts were subjected to IP with anti-PCAF. The obtained immunoprecipitates were used as a source of active PCAF and were used for in vitro cdk2 acetylation experiments. Autoradiography indicates cdk2 acetylation (top panel). A WB anti-cdk2 of the membrane used for autoradiography is shown on the bottom panel. Asterisks indicates non-specific bands. (D) The immunoprecipitates obtained in (C) were checked for the content of PCAF and SPT3 by WB.