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. 2022 Oct 11;30(3):716–730. doi: 10.1038/s41418-022-01074-0

Fig. 4. C9orf72 directly interacts with DNA-PKcs.

Fig. 4

a, b C9orf72 interacts with DNA-PKcs directly. a The purified Flag-C9orf72 protein pulled down DNA-PKcs in the cell lysate. Quantitative analysis of DNA-PKcs intensity. b HA-C9orf72 protein directly interacted with Flag-tagged KIP domain of DNA-PKcs in in-vitro pulldown assay. Data are mean (n = 3–5 independent experiments; paired t-test; *, p < 0.05). c, d DNA-PKcs deficiency impaired interaction between C9orf72 and Ku70, Ku80. HEK293T cells were transfected with Flag-C9orf72 and siDNA-PKcs (c), or siKu70 (d), and immunoprecipitated with anti-Flag antibody. The immunoprecipitated complex was probed with indicated antibodies. Quantitative analysis of intensities of Ku70, Ku80, and DNA-PKcs. Data are mean (n = 3 independent experiments; paired t-test; *, p < 0.05; **, p < 0.01). e, f The C terminus of C9orf72 is required for interaction with DNA-PKcs. e Schematic diagram of C9orf72 constructs. f HEK293T cells were transfected with C9orf72-Flag or indicated deletions. The immunoprecipitated complex was probed with DNA-PKcs antibody. Quantitative analysis of DNA-PKcs intensity. Data are mean (n = 4 independent experiments; paired t-test; *, p < 0.05; ***, p < 0.001).