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. 2013 Jul 1;94(Pt 7):1554–1567. doi: 10.1099/vir.0.051169-0

Fig. 6.

Fig. 6.

PEDV PLP2 interacts with and deubiquitinates RIG-I and STING. (a, b) PEDV PLP2 interacts with RIG-I and STING. HEK293T cells were transfected with Flag-tagged RIG-I (a) or STING (b) with V5-tagged PLP2, and co-immunoprecipitation experiments were performed with the indicated antibodies. The co-immunprecipitates were blotted with anti-V5 (top panel) and reprobed with anti-Flag (second panel) for RIG-I or STING detection. The input lysates were blotted with anti-V5 to detect PLP2 (third panel) and anti-Flag to detect RIG-I or STING (bottom panel). (c, d) PEDV PLP2 deubiquitinates RIG-I and STING in a catalytic-dependent manner. HEK293T cells were co-transfected with Flag-tagged RIG-I (c) or STING (d) together with HA-tagged Ub-K63, V5-tagged PLP2 or the catalytic mutants of C1729A, H1888A and D1901A. The lysates were immunoprecipitated with anti-Flag antibody and the immunoprecipitates were blotted with anti-HA to detect Ub-K63-conjugated RIG-I (c) or STING (d) (top panel) and reprobed with anti-Flag for RIG-I or STING detection (second panel). The input lysates were blotted with anti-V5 to detect PLP2 and the catalytic mutants (third panel) and anti-Flag to detect RIG-I and STING (bottom panel).