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Fig. 4 | Virology Journal

Fig. 4

From: Subcellular localization and interactions of Infectious Salmon Anemia Virus (ISAV) M1 and NEP as well as host Hsc70

Fig. 4

Co-localization and interaction of Hsc70-M1 with M1 or/and NEP. a Co-localization of Hsc70-M1 and Hsc70-NEP. SSN-1 cells were transfected with pEYFP-Hsc70 and pDsRed2-NEP, or pEYFP-Hsc70 and pEGFP-M1. At 24 h post-transfection, cells were fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100 and incubated DAPI. Cells were imaged on a LSM510 Meta confocal laser-scanning microscope. b Interaction of Hsc70-M1 and Hsc70-NEP. The His-M1 protein (10 μg) or His-Hsc70 (10 μg) was incubated with 10 μg GST, GST-Hsc70 or GST-NEP. Western blot analysis was performed using either anti-His (lower panel) or anti-GST (upper panel). In this assay, 1/100 of the total volume of His-M1 or His-Hsc70 was loaded as the input (lanes 1 and 2) and 1/10 of the total volume of each eluted sample was loaded to detect interacted proteins (lanes 3 and 4)

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