Fig. 1

Phylogenetic analysis of NEDD8 proteins and the expression profile of rabbit NEDD8 in different tissues. A, alignment of NEDD8 amino acid sequences from Homo sapiens (Hs), Oryctolagus cuniculus (Oc), and Danio rerio (Dr). B, phylogenetic tree of NEDD8 proteins in 11 species constructed using the neighbor-joining method. Homo sapiens (Hs), Gene ID: 4738; Oryctolagus cuniculus (Oc), Gene ID: 100009008; Mus musculus (Ms), Gene ID: 18002; Sus scrofa (Ss), Gene ID: 100520086; Gallus gallus (Gg), Gene ID: 100858776; Latimeria chalumnae (Lc), Gene ID: 102345978; Xenopus tropicalis (Xt), Gene ID: 549727; Danio rerio (Dr), Gene ID: 368667; Oryzias latipes (Ol), Gene ID: 101172934; Takifugu rubripes (Tr), Gene ID: 101077379; Drosophila melanogaster (Dm), Gene ID: 35151. C, predicted protein structure of rabbit NEDD8 by UniProtKB in the Ensemble database ( https://www.uniprot.org/uniprotkb/Q4PLJ0/entry ). D, mRNA expression levels of NEDD8 in various tissues of adult rabbits. Data are expressed as mean ± SD, n = 3 experiments. ?p < 0.05, ??p < 0.01, and ???p < 0.001. E, immunofluorescence staining of NEDD8 in follicles at different developmental stages in rabbit ovaries. The scale bar represents 20 ?m and 100 ?m, respectively. F, confocal microscopy image of endogenous NEDD8 localization in rabbit GCs. NEDD8 is stained red and nuclei are stained blue with DAPI. The scale bar represents 50 ?m. DAPI, 4?, 6-diamidino-2-phenylindole; EV, early vitellogenic stage; FG, full-grown stage; GC, granulosa cell; MV, mid-vitellogenic stage; ns, not significant; PG, primary growth stage; PV, previtellogenic stage.