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. 2020 Sep 10;12(9):1009. doi: 10.3390/v12091009

Figure 2.

Figure 2

Development of Mab recognizing endogenous swine ISG15. (A) Recombinant expressed sISG15 protein with serial dilutions were subjected to Western blotting using the antibody present in hybridoma culture supernatant (clone no. 3D5E6) to detect binding of Mab-3D5E6 to its immunogen. (B) Cell lysates from CRL-2843 treated with or without porcine IFN-α (pIFN-α) were harvested and subjected to Western blotting using the hybridoma culture supernatant (clone no. 3D5E6) to detect binding of Mab-3D5E6 to endogenous ISG15 in cell lines of swine origin. (C) CRL-2843 cells were either treated with pIFN-α or left untreated for 24 h. After cells were fixed and permeabilized, cells were incubated with Mab-3D5E6 followed by visualization after addition of Alexa Fluor® 555-labeled goat anti mouse IgG (H + L) (Red channel) and counter-staining with DAPI.