Phospho histone h2b ser14

MST2 (Abcam, ab52641), phospho‐histone H2B (Ser14) (Cell Signalling, 6959), phospho‐histone H2B (Ser14) (Millipore, 07‐191), MST1 (Millipore, 07‐061), RASSF1A (3F3, Santa Cruz sc‐58470), UBF (F9, Santa Cruz, sc‐13125), SAV‐1 (Atlas, HPA001808), V5 (Cell Signalling, 13202), H2B (abcam, ab52484), H2A (Abcam, ab18255), H3 (96C10, Cell Signalling, 3638), H4 (Cell Signalling, 2592), γH2AX (JBW301, Millipore, 16‐193), nucleolin (4E2, Abcam, ab13541), lamin A/C (Cell signalling, 4777), α‐tubulin (B3, Sigma, T9822), RCC1 (Cell Signalling, 5134), phospho‐MST1 (Thr183)/MST2(Thr180) (Cell Signalling, 3681), KAP1 (Bethyl, A300‐274A) and phospho‐KAP1S824 (Cell signalling, 4127).

Cells were grown on coverslips and treated as indicated. Cells were fixed with methanol at −20°C for 10 min, washed with 1× PBS and blocked with 2% BSA in 1× PBS. Coverslips were incubated with the indicated antibodies in blocking solution overnight at 4°C, washed and stained with secondary anti‐rabbit and or anti‐mouse IgG conjugated with Alexa Fluor secondary antibodies (Molecular Probes) for 1 h at room temperature. Coverslips were washed with PBS + 0.1% Tween, and DNA was stained with DAPI. Cells were analysed using LSM780 (Carl Zeiss Microscopy Ltd) confocal microscope. At least 200 cells were counted from three independent experiments. For detection of H2BS14p, the phospho‐histone H2B (Ser14; Cell Signalling, 6959) was used unless stated otherwise.