Figure 3 Competition binding experiments using the NPY Y 1 receptor-expressing SK-N-MC cell line.
Briefly, the density of expressed β-ARs on UROtsa cells was determined by saturation binding experiments using the nonselective β-AR antagonist I-CYP as the radiolabel (NEN Life Sciences, Boston, MA).
Moreover, although residual electron density was observed for t1G and t1C nucleotides, binding experiments using the same target RNAs show that these interactions do not contribute substantially to target affinity.
The whole cell binding experiments used the same method but without sonication and membrane preparation.
To further assess whether the p52 Bcl3 complex senses A/T-and G/C-centric κB sites differently, we carried out in vitro DNA-binding experiments using the electrophoretic mobility shift assay (EMSA).
To test the FREX performance under such conditions, we repeated the FRET binding experiment using FN3BN+I75A, P48, and SH2 in the presence of 10% fetal bovine serum.
To determine if the canonical Cdc5 Myb domains (R1 and R2) can individually interact with RNA, we repeated the RNA binding experiment using recombinant Cdc5-R1 and Cdc5-R2.
A modified Hill equation, I n o r m = M C h C h + K 1 / 2 h where K1/2 is the apparent binding constant, h is the Hill coefficient, M is the maximum instensity, and C is the concentration of Crm1 and RanQ69L, was fit to each binding experiment using sum-of-squares minimization implemented with the Solver tool in Microsoft Excel.
Hence, we performed a ligand binding experiment using 8 replicates for each concentration of CTB applied so as to obtain highly quantitative and information-rich binding data with very little reagent material required.
Binding experiments, using CHO cells expressing the human SP-specific NK1 receptor, demonstrated the functionality of all peptidylcyclodextrin derivatives, which exhibited IC50 values in a 10−9 10−7 M range.
