Selective Sorting of Senescent Cell Subpopulations Compatible with Downstream Genomics Applications

The dynamic character of senescence renders detection and selection of senescent cells challenging. One key feature of senescence is the alteration of chromatin features, and many methods for studying chromatin require only mild fixation of cells. The recent development of GLF16 compound allows for the selection of senescent cells in a population via fluorescence-activated cell sorting. Here, we detail two versions of a modified protocol that uses GLF16 to selectively sort senescent cells so as to be used in downstream genomics applications that require special fixation and permeabilization conditions. As proof of principle, we sort a subpopulation of senescent fetal lung fibroblasts and subject it to standard transcriptomics analysis, while the same procedure could potentially be coupled to other assays like CUT&RUN, CUT&Tag, ATAC-seq, or Hi-C/Micro-C.