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. 2021 Nov 11;297(6):101399. doi: 10.1016/j.jbc.2021.101399

Figure 2.

Figure 2

Verification of MS data with Western blot analysis shows the association of nsp1 with both nuclear pre-mRNA processing proteins and stress granule proteins.A, streptavidin pull down was conducted from cells expressing BioID2-fused nsp1 in the presence of 100 μM biotin followed by immunoblot of nuclear proteins involved in pre-mRNA cleavage and polyadenylation. B, streptavidin pull down and immunoblot of proteins involved in heat shock, stress, and translation from BioID2-fused nsp1-expressing cells grown in the presence of 100 μM biotin. C, analysis of the localization of nuclear proteins in the presence of nsp1. Either nsp1-Myc (SARS-CoV) or nsp1-BioID2 (SARS-CoV-2) were expressed in HEK cells for 24 h followed by the separation of nuclear and cytoplasmic fractions. Each fraction was analyzed in an immunoblot using antibodies against nuclear pre-mRNA processing proteins (NUDT21 and XRN2), cytoplasmic proteins (G3BP1 and PABPC1), and a control (GAPDH). HEK, human embryonic kidney cell line; nsp1, nonstructural protein 1; SARS-CoV, severe acute respiratory syndrome coronavirus; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.