Rabbit anti mx1 antibody

PC3 cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured with RPMI 1640 culture medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% v/v fetal bovine serum (FBS) (Internegocios, Buenos Aires, Argentina), penicillin 100 U/mL, streptomycin 100 µg/mL, and amphotericin 0.5 µg/mL. Hemin was obtained from SIGMA-Aldrich (St. Louis, MO, USA). For treatments, cells were incubated 24 h in RPMI media containing 10% FBS and antibiotics and then were exposed to Hemin (80 µM, 24 h). Thapsigargin was obtained from Sigma-Aldrich and dissolved in dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS). For treatments, cells were incubated in complete RPMI media and treated with Thapsigargin 0.1 µM or 0.25 µM for 24 h. Interferon gamma (INFγ) protein was obtained from ImmunoTools (Friesoythe, Germany). PC3 cells were treated with INFγ, 500 U/mL, for 18 or 24 h.
Mouse anti-human HO-1 monoclonal antibody and rabbit anti-MX1 antibody were obtained from Abcam (Cambridge, UK). Mouse anti-human β-actin antibody, rabbit anti-LC3, and rabbit anti-GADPH antibodies were obtained from Cell Signaling (Danvers, MA, USA). Horseradish peroxidase (HRP) conjugated anti-mouse secondary antibody was obtained from Cell Signaling. Secondary antibodies conjugated to Alexa 555 and Alexa 647 fluorophores were obtained from Molecular Probes, Invitrogen.

KU-CBE cells were infected with H3N2 CIV at 0.1 MOI or treated with 1 g/ml of canine IFN-β or IFN-α (Cloud-Clone Corp, USA) for 24 h. After cells were lysed using the RIPA buffer (Sigma-Aldrich), the MX1 and OAS1 proteins were detected in cell lysates using western blots as described above. The rabbit anti-OAS1 antibody (Abcam) and rabbit anti-MX1 antibody (Abcam) were used as primary antibodies.