Expression of USP5 eukaryotic plasmid in vitro and its subcellular localization. A) Expression of USP5 eukaryotic plasmid in vitro. B) Localization of USP5 in FHM cells. Transfected the USP5 plasmid and Vector into FHM cells. Indirect immunofluorescence was performed after 24 h. The nucleus of DAPI-stained cells was blue, and the green fluorescence was USP5 with HIS tag. Observe under a forty magnification. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

USP5 enhances the expression of RIG-I. A) USP5 enhances the protein level of RIG-I. Co-translate USP5 and Vector with RIG-I, respectively. Protein samples were collected after 24 h. Detection of changes in protein levels. B) USP5 enhanced RIG-I protein level in a dose-dependent manner. In FHM cells, USP5 eukaryotic expression plasmid 0ug, 0.5ug, 1ug, 1.5ug and Vector 1.5ug,1ug,0.5ug,0ug were co-transformed with 1ug RIG-I, respectively. After 24 h, the expression levels of RIG-I and USP5 were detected. C) USP5 can activate higher IFNφ1 levels stimulated by Poly (I: C). 1ug USP5 or 1ug Vector was transfected into FHM cells in the presence or absence of poly (I: C). Samples were collected after 24 h. Isolate RNA. qRT-PCR was used to detect the mRNA expression level of IFNφ1. N = 3, P value < 0.05 was marked with *, p < 0.01 was marked with **, p < 0.001 was marked with *** and p < 0.0001 was marked with ****.