Immunoblotting Protocol for Viral Protein Detection

For immunoblotting, cell lysates were mixed with 4X NuPAGE LDS sample buffer and 5% beta-mercaptoethanol. Lysates were resolved by SDS-PAGE and transferred to 0.45 μM PVDF membranes. For detection of protein, primary antibodies used were mouse anti-firefly luciferase (1:1000 dilution, MA1-12556, Invitrogen), mouse-anti PA (1:500 dilution, F5-32^{75 (link)}), mouse anti-PB1 (1:500 dilution, F5-10^{75 (link)}), mouse anti-beta actin (1:10,000 dilution, 8H10D10, Cell Signaling Technology), rabbit anti-Lamin A/C (1:1000 dilution, 2032, Cell Signaling Technology), mouse anti α-Tubulin (1:500 dilution, 2144, Cell Signaling Technology), and rabbit anti-GRSF1 (1:3000 dilution, A305-136A, Bethyl Laboratories). Secondary antibodies used were rabbit anti-mouse IgG HRP-linked (1:5000 dilution, 7076, Cell Signaling Technology), goat anti-rabbit IgG HRP-linked (1:5000 dilution, 7074 Cell Signaling Technology) and goat anti-mouse IgG1 HRP-linked (1:3000 dilution, ab97240, abcam).