LLPS of NONO promotes the association of EGFR and NONO. (A) FRAP of NONO-GFP droplets in vivo. (B) 1,6-Hexanediol disrupted NONO-GFP droplets. 5% 1,6-Hexanediol in complete medium was added at 0 second and removed at 30 seconds. (C) purified NONO-GFP protein formed LLPS in vitro. RNA extracted from HEK293T cells were added to a concentration of 200 ng/ul. (D) NONO droplets recruit EGFR and DNA-PK. The nuclear protein was fractionated from U2OS cells. (E, F) confirm the impact of 1,6-Hexanediol on EGFR/NONO and EGFR/DNA-PK complex via Duolink PLA assay. (G) 1,6-Hexanediol suppressed IR-induced autophosphorylation of DNA-PK at T2609. For (E-G), 2% 1,6-Hex was added after irradiation and cells were cultured for 15 (E, F) or 30 minutes. ***, P < 0.001.