<p>Grass carp NHRE3 degrades ISG15 via the K48-linked ubiquitination–proteasome pathway.</p>

(B) After overexpressing NHRE3, the CIK cells were infected with GCRV, WB was used to detect ISG15 protein expression after 24h, with the uninfected and empty vector overexpression groups serving as controls. (C) After overexpressing <i>nhre3</i>, the CIK cells were infected with GCRV, qPCR was used to detect isg15 gene expression after 24h, with the uninfected and empty vector overexpression groups serving as controls.

(D) 293T cells were co-transfected with 0, 1, or 2 μg of pcDNA-NHRE3-HA and pCMV-ISG15-MYC. After 18 h, the cells were treated with MG-132 (20 μM) for 6 h, and ISG15 degradation was assessed via western blot. (E) 293T cells were co-transfected with pcDNA-NHRE3-HA, pCMV-ISG15-MYC, and pCMV-Ub-His.