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We have utilized a similar approach with further modification to image the mitochondria and validated for large-scale compound screening.
Compound screening for potential antivirals and clinical isolate titrations were successfully arranged.
The PIK3Ca mutation status was also correlated with compound screening data for the cell lines.
This work, we believe, can minimize the cost of chemical compound screening for the biotechnological industry.
This approach biases the chemical profiles of compound screening collections towards known structures.
For compound screening and target identification measurements cells were treated with 200 µM BSO alone or in co-incubation with semi-logarithmic dilutions of the test compound.
A high-throughput image-based approach is also described for easy quantification of apoptosis and necrosis so that large-scale compound screening is also possible in a rapid manner.
Even though the above-described approach is sensitive to distinguish necrosis from apoptosis in real-time mode, for compound screening the approach needs to be adapted for high-throughput mode.
Nonetheless, the fact that this protein family is frequently associated with cancer, often with activating mutations, probably warrants investment into biological target validation, assay development and compound screening to identify inhibitors of these enzymes.
Utilizing tissue-relevant ECM coatings for neuronal cultures allows accelerated network formation and maturation within two weeks (rather than four weeks in the absence of ECM), thus providing the means to increase throughput of functional compound screening applications.
Although MHC-I alleles have been reported in cynomolgus macaques39,40, we did not determine their precise haplotypes in this study, in an effort to induce less bias regarding MHC haplotypes during our compound screening.
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