Anti lig3

Whole-cell extracts were collected in SDS sample buffer and boiled for 10 mins. Samples were resolved by SDS-PAGE, transferred to PVDF, and blocked with 5% milk in PBST (PBS, 0.1% Tween-20). The following primary antibodies were used at 1:1,000 dilution (unless noted) in PBST: anti-CENP-A (Cell Signaling, 2186), anti-phospho histone H2AX (ser139) clone JBW301 (EMD Millipore, 05-636), anti-phospho histone H3 (ser10) (Cell Signaling, 9706), 1:4,000 anti-histone H3 (Sigma H0164), anti-LIG4 (GeneTex, GTX100100), anti-DNA-PKcs (Bethyl, A300-516A), anti-LIG3 (Bethyl, A301-637A), anti-PARP (BD Pharmingen, 556362, kindly provided by X. Wu, The Scripps Research Institute, USA), anti-BRCA2 (Bethyl, A303-434A), anti-RAD51 (Abgent, AM8421b), and 1:2,000 anti-GAPDH (Cell Signaling, 14C10). Blots were probed with 1:4,000 dilutions of HRP-conjugated secondary antibodies (GE Healthcare) and exposed to film. All unprocessed film scans with the appropriate size markers are provided in Supplementary Fig. 6.

The following antibodies were used: anti-TIMELESS (Bethyl Laboratories, A300-961A and A300-960A, the latter used for IP), anti-PARP1 (Invitrogen, 436400), anti-PARP1 (Cell Signaling Technology, 9542 and 46D11, the latter used for IF), anti-FLAG M2 (Sigma, F3165), anti-p-Chk1 (Cell Signaling, 2344), anti-p-Chk2 (Cell Signaling, 2661), anti-p-ATM (Cell Signaling, 4526), anti-LIG3 (Bethyl, A301-637A), anti-HLTF (Bethyl, A300-229A), anti-KU70 (Santa Cruz Biotechnology, sc-9033), anti-KU80 (Neomarkers, MS-285-P1; Cell Signaling, 2180), anti-XRCC1 (Cell Signaling, 2735), anti-DNA-PK (Santa Cruz, sc-5282), anti-DNA-PK (Cell Signaling, 12311), anti-TIPIN (Bethyl, A301-474A-1), anti-RPA1 (Santa Cruz, sc46504), anti-RPA2 (Millipore, 04-1481), anti-SSRP1 (Abcam, ab26212), anti-SPT16 (Cell Signaling, 12191), anti-SKP1 (Santa Cruz, sc-5281), and anti-phospho-H2AX (Millipore, 05-636).