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. 2006 Dec 11;27(4):1296–1308. doi: 10.1128/MCB.00336-06

FIG. 3.

FIG. 3.

Two residues, K17 and R18, of SRC-3 essential for its protein turnover rate. (A) FLAG-SRC-3 wt (FLAG-SRC3) or its double mutant (K17A/R18A mutant), along with β-Gal, was expressed in 293 cells. After treatment with CHX for the indicated times, cells were harvested and analyzed by Western blotting using antibodies as indicated. PARP, poly(ADP-ribose) polymerase. (B) Pulse-chase analysis of SRC-3 wt and its K17A/R18A mutant. Inducible 293 cells for overexpressing SRC-3 wt or the K17A/R18A mutant were induced for 2 days, followed by 35S labeling and chasing for times indicated. 35S-labeled SRC-3 protein was analyzed by IP with anti-FLAG antibodies, followed by (a) SDS-PAGE and (b) phosphorimager quantitation.