Figure 8 | Scientific Reports

Figure 8

From: Interaction of PIAS1 with PRRS virus nucleocapsid protein mediates NF-κB activation and triggers proinflammatory mediators during viral infection

Figure 8

Modification of N by SUMO2/3 conjugation. (a) MARC-145 cells were infected with PRRSV at 5 moi for 48 h, and cell lysates were subjected to co-IP using α-PRRSV-N MAb (MR40) for immunoprecipitation and α-PRRSV-N PAb, α-SUMO1 MAb, or α-SUMO2/3 PAb for immunoblot (upper panels). Whole cell lysates (WCL) were subjected to Western blot as an input control (bottom panels). (b) HeLa cells were transfected with FLAG-N plasmid (2 μg) along with the HA-SUMO1 (2 μg), HA-SUMO2 (2 μg), or HA-SUMO3 (2 μg) plasmids. After 24 h, cells were stained with α-FLAG PAb (red) and α-HA MAb (green) to detect N and SUMO, respectively. Nuclei were stained with DAPI (blue). (c) HeLa cells were transfected with FLAG-N plasmid (2 μg) for 30 h, and cell lysates were subjected to co-IP using α-FLAG MAb for immunoprecipitation and α-SUMO2/3 PAb or α-FLAG PAb for immunoblot. Whole cell lysates (WCL) were subjected to Western blot as an input control (bottom panels), and β-actin served as a loading control.

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