Dub sirna library on targetplus

The DUB siRNA library (ON-TARGETplus) and PARP-1 siRNA containing pools of 4 individual siRNAs and an individual siRNA targeting USP6 (USP6_13 5′-CAGCUAAGAUCUCAAGUCA-3′) were from Horizon Discovery (Cambridge, UK). The non-targeting control siRNA (AllStars Negative Control siRNA) was from Qiagen (Manchester, UK). The following antibodies were used: USP6 and PARP-1 (both Santa Cruz Biotechnology, Heidelberg, Germany); phospho(T68)-Chk2 (New England Biolabs, Hitchin, UK); XRCC1 and APE1 (kindly provided by G. Dianov); γH2AX (Merck-Millipore, Watford, UK); 53BP1 (Bethyl Labs, Montgomery, AL); and actin (Sigma-Aldrich, Gillingham, UK).

The DUB siRNA library (ON-TARGETplus), USP9X siRNA pool and individual siRNA targeting USP9X (USP9X_6 5′-AGAAAUCGCUGGUAUAAAU-3′ and USP9X_8 5′-GUACGACGAUGUAUUCUCA-3′) were from Horizon Discovery (Cambridge, UK). The non-targeting control siRNA (AllStars Negative Control siRNA) was from Qiagen (Manchester, UK). The following antibodies were used: PARP-1 (sc-53643) and Mcl-1 (sc-819; both Santa Cruz Biotechnology, Heidelberg, Germany), γH2AX (05-636; Merck-Millipore, Watford, UK), USP9X (A301-350A) and 53BP1 (A300-272A; both Bethyl Labs, Montgomery, AL), LC3B (2775) and β-Galactosidase (9860; both Cell Signaling Technology, London, UK), Pericentrin (ab4448; Abcam, Cambridge, UK), CEP55 (23891-1-AP) and CEP131 (25735-1-AP; both Proteintech, Manchester, UK) and tubulin (T6199; Sigma-Aldrich, Gillingham, UK). Goat anti-mouse Alexa Fluor 555 (A21422) or goat anti-rabbit Alexa Fluor 488 (A11008) secondary antibodies for immunofluorescence were from Life Technologies (Paisley, UK).