Skip to main content
. 2016 Jun 9;7(34):54430–54444. doi: 10.18632/oncotarget.9914

Figure 1. Crosstalk between BER and DSB repair after irradiation.

Figure 1

A. Loss of APE1 or NEIL1 causes moderate radiosensitization. For clonogenic survival analysis (left panel), HEK293 cells were irradiated at 48 h after transfection with siRNAs. Immunoblotting (right panel) shows depletion of target proteins. B. The FLAG-NEIL1 co-IP in HEK293 cells revealed radiation-associated increase in association of Ku, DNA-PKcs, and PNKP but not of XRCC4 with NEIL1. C. The IP of WT APE1 but not of the NΔ33 mutant (lacking APE1's common interaction domain) contained Ku and BER/SSBR proteins, The FLAG IPs were isolated from HEK293 cells after transfection with FLAG-tagged WT APE1 or the NΔ33 mutant. D. The FLAG-NEIL1/2 IP contained DNA-PKcs and Ku, but the IP of NEIL1(N311) mutant that lacks the common interaction domain (aa312-389) did not contain DNA-PKcs or Ku, underscoring the specificity of Ku interaction. E. The FLAG-Ku IP contained XRCC4, APE1, and NEIL1, whose levels increased after bleomycin treatment. F. Endogenous NEIL1 co-IP contains Ku70, DNA-PKcs after irradiation, unlike PNKP and XRCC1, which are constitutively associated. G. PLA analysis confirms in-cell association of NEIL1 with Ku, which was enhanced after IR treatment (> 25 cells were counted for the bar graph).