IPed samples were prepared as previously described (27 (link)). Briefly, HEK293T cells were transiently transfected with SFB-tagged ZMYM2. Cells were extracted with NETN buffer for 30 min at 4°C. Cell lysates were centrifuged, and the supernatants were collected as the soluble fraction. The pellets were digested with NETN buffer with Turbo-Nuclease and 1 mM MgCl2 for 1 h at 4°C and collected as the chromatin fraction. The both fractions were incubated with streptavidin sepharose bead (GE healthcare) for 1 h at 4°C. The beads were washed twice with NETN buffer and eluted with 2 mM biotin at 4°C. The eluted supernatants were incubated with S-protein beads (EMD Millipore) overnight at 4°C and washed three times with NETN buffer. The protein mixtures were eluted by boiling with 1× sample buffer. The eluted complex was subjected to SDS-PAGE and MS analysis.
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