ChIP-seq of STAT5B in human ALCL cell lines and CDK6 in a murine ALCL cell line
ChIP-seq of STAT5B in human ALCL cell lines and CDK6 in a murine ALCL cell line
A total of 1.5 × 10e7 cells per sample were collected from cell culture and washed twice with ice-cold PBS containing 1 mM PMSF. Cells were crosslinked with 2 mM disuccinimidyl glutarate (DSG) in PBS under rotation at room temperature for 30 minutes. Cells were then washed and incubated with 1% formaldehyde in PBS for 10 minutes at room temperature under rotation. The formaldehyde crosslinking was quenched by adding 0.125 M glycine and rotating for 5 minutes. For immunoprecipitation, the following antibodies were used:  Anti-STAT5B (5 µg; [EPR16671], Abcam, ab178941),  Anti-CDK6 (5 µg; rabbit polyclonal, PA5-27978; Invitrogen, USA).  Input DNA or DNA enriched through IP was subjected to sequencing library preparation using the NEBNext Ultra II chemistry (New England Biolabs) 