Ab47476

Lipopolysaccharide (LPS) and SP600125 were purchased from Sigma-Aldrich (St Louis, MO, U.S.A.). The cell culture Medium (DMEM, MEM, and RPMI 1640), fetal bovine serum (FBS), penicillin–streptomycin (10,000 U/ml) solution, and phosphate-buffered saline (PBS) were purchased from Thermo Fisher Scientific (Waltham, MA, U.S.A.).
Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α (MTA00B), IL-6 (M6000B), and IL-1β (MLB00C) were obtained from R&D Systems (Minneapolis, MN, U.S.A.). The primary antibodies for Ufm1 (ab109305), LZAP (CDK5RAP3) (ab157203), NF-κB p65 (ab16502), p-NF-κB p65 (ab86299), ATF2 (ab47476), p-ATF2 (ab32019), c-Jun (ab32137), p-c-Jun (ab32385), anti-GFP antibody (ab290), and β-actin (ab8226) were from Abcam (Cambridge, U.K.). Anti-Flag antibody (F7425) was form Sigma. Anti-mouse IgG horseradish peroxidase-conjugated secondary antibody (ab97040) was also from Abcam.

Proteins were extracted using lysis buffer. Protein concentration was assessed by BCA method (Beyotime). Total proteins was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membranes (Millipore, USA), then blocked with 5% skim milk. After blocking, the membranes were incubated with primary antibodies followed by incubation with secondary antibodies. The signal was detected by chemiluminescence (Thermo Fisher Scientific). Primary antibodies against MARCH6 (ab183533) and ATF2 (ab47476) were purchased from Abcam. GAPDH (60004–1-Ig) antibody was purchased from Proteintech.