H4K77ac RGFP966 AgedHSC rep2
CUT&Tag assays were performed by using a NovoNGS CUT&Tag 3.0 High-Sensitivity Kit per the manufacturers instructions (Novoprotein Scientific). In brief, FACS-sorted HSCs from young or aged mice (20,000 cells for each) were suspended in primary antibody buffer with 0.25 g of anti-H4K77ac antibody and incubated overnight at 4 C, followed by incubation with a secondary antibody (guinea pig anti-rabbit secondary antibody, ABIN101961) and the pA-Tn5 adapter complex. The libraries were then amplified by mixing DNA with barcoded i5 and i7 primers (see Supplementary Table 7 for details), followed by PCR amplification. For sequencing, the libraries were assessed for quality and quantity using the Agilent 2100 Bioanalyzer (Agilent Technologies) and Qubit fluorometer (Invitrogen) and sequenced on the Illumina X10 Sequencer in 150-bp paired-end format.