Fig. 3: cGAS promotes USP7-mediated DNA-PKcs deubiquitination and stabilization.
From: The cGAS-Ku80 complex regulates the balance between two end joining subpathways
A Western blotting analysis of the DNA-PKcs protein level in HEK293T cells transfected with the USP5, USP10 or USP36 overexpression vector. B Western blotting analysis of the DNA-PKcs protein level in cells transfected with the USP7 or enzymatically dead USP7 (USP7m) overexpression vector. C The expression of DNA-PKcs in HEK293T cells with USP7 depletion. D, E Co-IP analysis of the interaction between endogenous USP7 and DNA-PKcs in HEK293T cells using antibodies against USP7 or DNA-PKcs. F In vitro co-IP analysis of the interaction between USP7 and DNA-PKcs. The two purified recombinant proteins were incubated, and a co-IP assay with an antibody against GST was performed, followed by Western blotting analysis. CBB, Coomassie Brilliant Blue. G Analysis of the level of K48-linked ubiquitination of DNA-PKcs in USP7-depleted HEK293T cells. Cells were treated with MG132 (10 μM, 12 h) before being harvested for co-IP and Western blotting analysis. H Co-IP analysis of the interaction between USP7 and DNA-PKcs in HEK293T cells overexpressing cGAS-HA. I Western blotting analysis of the DNA-PKcs protein level in control and USP7-depleted HEK293T cells with or without cGAS overexpression.
