Anti xrcc5

4×10⁴ cells per well were cultured in 24-well plates. After adherence, cells were fixed by 4% paraformaldehyde for 15 min at room temperature. After washed with PBS, cells were blocked in 5% normal goat serum with 0.3% Triton-X100 for 30 min. Cells were incubated with a mixture of primary antibodies for overnight at 4℃ after washed with PBS. The primary antibodies used were as follows: mouse monoclonal anti-DTX3 (1:100, cat. no. sc-376439, Santa Cruz Biotechnology, Inc) and rabbit polyclonal anti-XRCC5 (1:200, cat. no. 16389-1-AP, Proteintech Group Inc). Then cells were washed with PBS 3 times for 5 min and incubated with a mixture of secondary antibodies, goat anti-mouse IgG Dylight 594 (1:100) and goat anti-rabbit IgG Dylight 488 (1:100), for 1 h. For nuclei staining, cells were counterstained with DAPI (cat. no. C0065, Solarbio Life Sciences) for 5 min and mounted with Antifading Mounting Medium (cat. no. S2100, Solarbio Life Sciences) for confocal image acquisition (400× magnification).

Etoposide, MMS, TSA, NIM, and thymidine were purchased from Sigma-Aldrich. Hydrogen peroxide (H₂O₂) was purchased from Duksan Pure Chemicals (Ansan, Korea). The following antibodies were used in this study: anti-XRCC5 (sc-5280; Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:1000), anti-XRCC6 (sc-55505; Santa Cruz Biotechnology; 1:1000), anti-β-actin (sc-47778; Santa Cruz Biotechnology; 1:3000), anti-γ-H2AX (#9718S; Cell Signaling Technology, Danvers, MA, USA; 1:1000), anti-PARP-1 (sc-74469; Santa Cruz Biotechnology; 1:1000), anti-α-tubulin (LF-PA0146; AbFrontier, Seoul, Korea; 1:1000), anti-SIRT1 (sc-74504; Santa Cruz Biotechnology; 1:1000), anti-Myc (#2276 S; Cell Signaling Technology; 1:1000), anti-Flag (#2368S; Cell Signaling Technology; 1:1000), anti-HA (sc-805; Santa Cruz Biotechnology; 1:1000), anti-His (sc-803; Santa Cruz Biotechnology; 1:1000), anti-acetylated lysine (sc-32268; Santa Cruz Biotechnology; 1:1000), and anti-phosphorylated serine (sc-81514; Santa Cruz Biotechnology; 1:1000). Polyclonal anti-FOXL2 antibodies (1:1000) were generated using either recombinant human FOXL2 protein or N-terminus peptides as an antigen^{51 (link)}. Other reagents were purchased from Sigma-Aldrich, unless otherwise indicated. Antibodies were validated for the specific species and application in pilot studies.