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The extent of potentiation at a fixed analogue concentration (10 μ M) was related to the potency of inhibition of thymidine uptake.
The observed rate constant (kobs) is then plotted versus the corresponding analogue concentration, and the data are fit to the rectangular hyperbola kobs = kpol[dNTP]/ Kd + [dNTP]) to give kpol and Kd parameters (Supporting Information Tables S1 and S2).
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The analogue concentrations used were based on the DHEA concentrations of serum levels found in humans and other species and were chosen to approximate analogue levels in vivo.
These results suggested that the analogue can bind to the putrescine-binding site and that, in the presence of putrescine or at high analogue concentrations, binding is shifted to that of the active site.
For induction assays, vitamin D3 analogues (concentrations ranging from 10-9 to 10-5 M) were added after 24 h and left for additional 48 h.
This FRET assay described above was used to examine the rate of reaction for all three mutant enzymes at high coumarin analogue (4) concentrations (data not shown).
These analogues induce concentration-dependent degradation of Hsp90-dependent client proteins with the six-membered bicyclic analogues manifesting increased activity versus the five-membered counterparts.
Cells were exposed for 1, 2 and 4 days, or for 6, 12 and 24 h in the case of cycle-synchronised cells, to staurosporine analogues at concentrations at which they inhibited growth by 80% after 4 day exposure.
Binding profile of MIPs and the corresponding NIPs in the presence of glucose and its analogue, fructose, at concentration ranges from 10to9090 mg l−1 is shown in Figure 4.
After 24-hour incubation to allow cells to adhere, cells were treated with silibinin (Sigma Aldrich), esculetin (Sigma Aldrich), curcumin (Sigma Aldrich), or vitamin D analog (unpublished new analogue) at different concentration (0.1, 1, 10, 100 μM).
Since all 177Lu analogues were obtained in significantly lower specific activities than the 68Ga analogues, the ligand concentration had to be increased to 0.5 nM to obtain reasonable count rates for assay evaluation.
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