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. Author manuscript; available in PMC: 2010 Aug 14.
Published in final edited form as: Mol Cell. 2009 Aug 14;35(3):352–364. doi: 10.1016/j.molcel.2009.06.015

Figure 3. Gcn5 loss alters TRF1 and POT1a protein turnover.

Figure 3

(A) Treatment of the cells with the proteasome inhibitor MG132 stabilizes TRF1 protein levels in Gcn5 null cells (lanes 2 and 4). Lysates from Gcn5 null and Gcn5 heterozygous MEFs, treated with MG132 or DMSO only, were resolved by SDS-PAGE, and TRF1 protein levels were monitored by immunoblot. (B and C) Decreased levels of exogenous FLAG-mTRFl and POT1a-Myc in Gcn5 depleted cells. MEFs with the indicated genotypes were transfected with mTRFl-FLAG expressing vector or infected with POT1a-Myc expressing retroviral vectors. After MG132 or DMSO treatment, lysates were prepared and the exogenous TRF1-FLAG levels were monitored by using anti-FLAG antibody. For monitoring POT1a-Myc protein levels, anti-Myc IP was performed. (D) Depletion of Usp22 has a similar impact on TRF1 protein levels as does Gcn5 depletion (compare lanes 1 and 2 to lanes5 and 6). (E) TRF1 co-immunoprecipitates with GCN5, USP22 and ATXN7L3.