Rabbit anti stat1

Immunoblotting analysis was performed as described in reference 23 (link). Briefly, approximately 2.10⁶ RAW264.7 cells were plated onto 35-mm dishes and infected the next day with MNV at an MOI of 10 as described above. At the indicated times, cells were lysed in 150 μl of 1× gel loading buffer (New England BioLabs), sonicated, and boiled 5 min at 95°C. Cell lysates were separated by SDS-PAGE using 10 μg of total proteins, and the proteins were transferred to polyvinylidene difluoride membranes. These were then probed with the following primary antibodies: mouse anti-phospho-ATF2 T71 (1:1,000; Abcam), rabbit anti-ATF3 (1:500; Cell Signaling), goat anti-TNF-α (1:500; Santa Cruz), rabbit anti-Ddx58 (1:500; Santa Cruz; sc-376845), rabbit anti-Stat1 (1:1,000; Abcam; Ab92506), rabbit anti-ATF2 (1:1,000; Cell Signaling; 35031), mouse anti-VP1 (1:500; I. G. Goodfellow, Cambridge, UK), anti-MNV-3 mouse immune sera (1:1,000; I. G. Goodfellow, Cambridge, UK), rabbit anti-NS7 (1:10,000; I. G. Goodfellow, Cambridge, UK), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Clone 6C5, 1:20,000; Invitrogen), followed by incubation with the appropriate peroxidase-labeled secondary antibodies (Dako) and chemiluminescence development using the Clarity Western ECL Substrate (Bio-Rad). The results were acquired using the Vilber imaging system.