Rabbit anti atf3

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.

Sangivamycin (HY-118384), rapamycin (HY-10219) and torin-1 (HY-13003) were obtained from MedChemExpress (NJ, USA). The primary antibodies used for western blotting were as follows: rabbit anti-ATF3 (#33593), rabbit anti-ATF4 (#118115) and rabbit anti-TRIB3 (#43043) were obtained from Cell signaling (MA, USA); rabbit anti-DDIT4 (#10638-1-AP) was obtained from Proteintech; mouse anti-actin (#sc-47778) was obtained from Santa Cruz Biotechnology (TX, USA). The ATF3 antibody (#18665) used for co-immunoprecipitation was obtained from Cell signaling (MA, USA). The primary antibodies used for immunochemistry staining: ki67 (#M7240, Agilent Technologies, CA, USA), p27 (#M72031, Agilent Technologies, CA, USA) and ATF3 (#MA5-31360, Thermo Scientific, MA, USA).