Anti mus81

Tissues and cells were lysed in RIPA buffer (cat#V900854, Sigma, MO, USA) and protein concentration was measured with the BCA method (cat#P0012, Beyotime, Shanghai, China). Proteins were isolated by SDS-PAGE, transferred to PVDF membranes (Millipore, Billerica, USA), incubated with primary and secondary antibodies and imaged with the ECL detection reagent (cat#12630, CST, California, USA) using a ChemiDoc™ XRS+ System. The primary antibodies included anti-Mus81 (1:1,000; cat#ab14387, Abcam, Cambridge, UK), anti-BRD4 (1:1,000; cat#ab128874, Abcam), anti-ZEB1 (1:1,000; cat#ab180905, Abcam), anti-E-cadherin (1:1,000; cat#3195, CST, California, USA), anti-N-cadherin (1:1,000; cat#13116, CST), anti-Snail1 (1:1,000; cat#9782, CST), anti-ZEB2 (1:1,000; cat#14026-1-AP, Proteintech, Wuhan, China), anti-Sirt5 (1:500; cat#15122-1-AP, Proteintech), anti-GAPDH (1:5,000; cat#G9545, Sigma). The secondary antibodies included HRP conjugated goat anti-Rabbit (1:3,000; SA00001-15, Proteintech) and anti-Mouse (1:3000; cat# SA00001-1, Proteintech).

Cell extracts were obtained by lysing cells for 15 min at 4C in buffer containing 50 mM Tris-HCl (pH 8.0), 300 mM NaCl, 0.4% NP-40, 10 mM MgCl2 and 5 mM DTT, supplemented with protease and phosphatase inhibitors (Halt Protease & Phosphatase Inhibitor Cocktail; ThermoFisher Scientific). After centrifugation (10,000 x g, 20 min), supernatants were diluted (v/v) in 50 mM Tris-HCl (pH 8.0), 0.4% NP-40 and 5 mM DTT. Proteins were separated by SDS-PAGE and immunoblotted with the following antibodies at dilutions recommended by the manufacturer: anti-pan-actin (MAB1501; Millipore), anti-ARTEMIS (#13381; Cell Signaling Technology), antimCherry (GTX128509; GeneTex), anti-CtIP (A300–488A; Bethyl), anti-FEN1 (A300–255A; Bethyl), anti-KAP1 (A300–274A; Bethyl), anti-DNA ligase3 (A301–636A; Bethyl), anti-MRE11 (A303–998A; Bethyl), anti-MUS81 (ab14387; Abcam), anti-PNKP (A300–257A; Bethyl), anti-SETX (A301–104A; Bethyl), anti-TDP1 (H00055775-A01; Abnova Corporation), anti-atubulin (T5168; Sigma-Aldrich), anti-XPF (A301–315A; Bethyl), anti-XPG (A301–484A; Bethyl), and anti-XRCC1 (A300–065A; Bethyl). Immunoblotting was revealed by chemiluminescence using a ChemiDoc MP Imaging System (Bio-Rad).