Skip to main content
. 2021 Jun 25;297(2):100921. doi: 10.1016/j.jbc.2021.100921

Figure 1.

Figure 1

Mapping the region of LigIII that interacts with TDP1.A, domain architecture of human XRCC1, DNA Ligase III, and TDP1. B, Upper panel, input (10%) of full-length and different LigIII domains. Lanes 1 to 6 represent LigIIIβ full length (1–846), LigIII-Ntase-OBD (387–846), LigIII-DBD (152–390), LigIII-ΔZnF (170–846), LigIII-ZnF-DBD (1–390), and LigIII-ZnF (1–136), respectively. Immunoblotting with mouse anti-6His mAb (Abcam). Lower panel, binding of Flag-TDP1 (10 pmol) to Ni-beads liganded by LigIII domains (10 pmol of each). Lane (∗) represents 10% TDP1 input. Negative control reaction lane, Ni-NTA beads alone with Flag-TDP1. Immunoblotting with mouse anti-Flag mAb (Sigma M2). C, Upper panel, input (10%) of GST-TDP1 and Flag-TDP1. Ni-NTA beads liganded by LigIIIβ (10 pmol) were used to pull down full-length Flag-TDP1, GST-TDP1, and GST-TDP1149–608 (10 pmol of each). The control lane (Control) represents Ni-NTA beads alone incubated with GST-TDP1. Immunoblotting with rabbit anti-TDP1-C-term (Abcam).