Co-crystallization with the nanobody NbGAK_4 trapped GAK in a dimeric arrangement similar to the one observed in the apo structure, whereas NbGAK_1 captured the activation segment of monomeric GAK in a well-ordered conformation, representing features of the active kinase.
The relatively large variability across different sessions within the same subject suggests multiple sessions should be used to accurately capture the activation patterns evoked by acupuncture stimulation at a particular point for a specific subject.
Thus using this construct, we could not capture the activation stage of PERK.
For each of the nine trial types, we defined one regressor per imaging run that captured the activation for each voxel across the three sequence executions.
To capture the activation of interarea circuits, it is necessary to investigate the dynamic spatiotemporal patterns with which excitation emerges in different cortical areas and, especially, their stimulus specificity.
As we can see on Figure 2, CNORfuzzy accurately captures the partial activation of p38, as well as the activation of ERK and, to some extent, the activation of NF κB.
It captures both the activation barrier (6.4 kcal mol−1) and the binding energy (−5.1 kcal mol−1) to good accuracy, as well as other properties such as weak interactions and hydrogen bonding (see Table S1 in the Supporting Information).
Because it only considers one time point, the model is also unable to capture the transient activation of ERK.
Nevertheless, the simulations are still unable to capture the prolonged activation of ATF4, because there is no other regulators that connect to ATF4 in the model to support its sustained activation after 6 h.
If, however, we wish to capture the transient activation of ERK, we can do so using a previously unpublished modification of the Boolean steady-state method which is available in CellNOptR.
Because the assay was well tolerated by cells and the assay chemistry is compatible with many different multiplexes, including other luminescent assays without the need for spectral filters, we were able to establish the optimal time to multiplex a luminescent apoptosis assay to capture the caspase activation window and avoid false-negative results.
