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Figure 1 | Veterinary Research

Figure 1

From: The tail domain of PRRSV NSP2 plays a key role in aggrephagy by interacting with 14-3-3ε

Figure 1

NSP2 induces autophagy. A Western blot analyses of the effect of NSP2 expression on LC3-II, LC3-I, and p62. 293T and BHK21 cells were transfected with empty vector, GFP-NSP2, or FLAG-NSP2. At 24 h after transfection, cell lysates were subjected to western blotting to analyze autophagy using anti-LC3 and p62 antibodies. The expression of NSP2 was verified by western blotting using anti-GFP or anti-FLAG antibodies. GAPDH served as the protein loading control. B Double-membrane phagophores structures were observed using an electron microscope. 293T cells were transfected with empty vector and GFP-NSP2 for 24 h. The cells were then fixed and sectioned, and the sections were observed using a transmission electron microscope. The red arrowheads indicate the double-membrane structure of autophagosomes. Co-localization of NSP2 and LC3 in 293T cells (C) and Marc-145 cells (D). 293T or Marc-145 cells were transfected with empty vector, GFP-NSP2, and GFP-NSP3 and were analyzed via confocal microscopy using anti-LC3B antibodies. EGFP-tagged proteins are green and LC3 proteins are red. Yellow represents the co-localization between proteins in the merged images. Cells transfected with the GFP vector served as negative control. Bar size: 25 µm.

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