Imaging-Based High-Content Screening with Clickable Probes Identifies XPB Inhibitors.

ABSTRACT: High-content screening (HCS) has become a powerful tool in drug discovery; however, its reliance on indirect readouts and surrogate markers limits HCS's ability to directly assess drug-protein interactions at endogenous levels, particularly in subcellular contexts. Here, we report an approach to address these limitations by combining confocal imaging-based HCS and bio-orthogonal labeling with clickable probes. As a proof-of-concept, we synthesized a probe Triptolide-alkyne (TL-alk) that rapidly and specifically labels xeroderma pigmentosum type B (XPB), a critical protein in nucleotide excision repair (NER). Probe-labeled XPB was conjugated to TAMRA to visualize the occupation of active sites, and EGFP and DAPI signals indicated XPB expression in the nucleus. Such a colorimetric HCS assay

SUBMITTER: Li S 

PROVIDER: S-EPMC12402854 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature