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. Author manuscript; available in PMC: 2011 Mar 23.
Published in final edited form as: Biochemistry. 2010 Mar 23;49(11):2475–2482. doi: 10.1021/bi901457e

Figure 1.

Figure 1

Intracellular stability of wild type and mutants with substitutions in Val3 of hTS. Stably transfected cells were treated with cycloheximide at the indicated time points and TS levels were analyzed by immunoblotting using an anti-human TS monoclonal antibody (D3B31). Blots were stripped and re-probed with actin as an internal control to normalize quantitation using ImageJ software. Half-lives were determined from the shown log2 plots of hTS levels (normalized against the actin content in each lane) determined by densitometry. Top panel: Western blot analysis of wt and Val3 mutant hTS protein levels after treatment with cycoheximide (CHX) for 0, 6,12,24,36, and 48 hours; Middle panel: Western blot analyses of V3Y, V3L, and V3F mutants after treatment with CHX for 0,2,4,6,8 and12 hours; Bottom panel: Log2 plot of the relative amount of protein in the band for hTS, (each unit corresponds to one half-life) for wt and Val3 mutants after treatment with CHX at the indicated time points.