Research Paper Volume 13, Issue 7 pp 9874—9899

DNASE1L3 arrests tumor angiogenesis by impairing the senescence-associated secretory phenotype in response to stress

DNASE1L3 interacts with H2BE. (A, B) Immunoblot analysis of DNASE1L3 in the subcellular localization under DNA damage using nucleocytoplasmic separation in two cell lines. (C) The result of blotting confirmed that DNASE1L3 translocated to the nucleus in response to DDR activation. (D) Lysates of 293T cells overexpressing Flag-H2BE and/or cMyc-DNASE1L3 were subjected to reciprocal co-immunoprecipitation (co-IP) to detect protein interaction. (E) HepG2 and HCCLM3 cell lysates were subjected to co-IP and immunoblot to detect endogenous H2BE and DNASE1L3 interaction. (F) FRET assay for DNASE1L3-H2BE interactions in living cells. FRET efficiency were calculated by Leica TCS SP8 software (FRETeff= (Dpost-Dpre)/Dpost). White circles identify FRET area. (G) Co-IP of cMyc-DNASE1L3 and Flag-tagged H2BE or H2BEΔ with N-terminal region (amino acids 1-28) deleted.

Figure 5. DNASE1L3 interacts with H2BE. (A, B) Immunoblot analysis of DNASE1L3 in the subcellular localization under DNA damage using nucleocytoplasmic separation in two cell lines. (C) The result of blotting confirmed that DNASE1L3 translocated to the nucleus in response to DDR activation. (D) Lysates of 293T cells overexpressing Flag-H2BE and/or cMyc-DNASE1L3 were subjected to reciprocal co-immunoprecipitation (co-IP) to detect protein interaction. (E) HepG2 and HCCLM3 cell lysates were subjected to co-IP and immunoblot to detect endogenous H2BE and DNASE1L3 interaction. (F) FRET assay for DNASE1L3-H2BE interactions in living cells. FRET efficiency were calculated by Leica TCS SP8 software (FRETeff= (Dpost-Dpre)/Dpost). White circles identify FRET area. (G) Co-IP of cMyc-DNASE1L3 and Flag-tagged H2BE or H2BEΔ with N-terminal region (amino acids 1-28) deleted.