Nf κb p65 rabbit polyclonal antibody

Western blotting was conducted as previously described [8 (link)]. Similarly, we obtained the supernatant of the sample and extracted proteins using the radio immunoprecipitation assay (RIPA) buffer containing 1 mmol/L PMSF (Beyotime, Shanghai, China), and the protein concentration was quantified using a BCA assay kit (Beyotime, Shanghai, China). During the process of electrophoresis, the proteins with different sizes were separated in SDS-PAGE. Then, the proteins on the gel were transferred onto a polyvinylidene-difluoride (PVDF) membrane (Beyotime, Shanghai, China) for blotting. Antibodies were acquired from Beyotime Biotechnology, Shanghai, China, which included glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (catalog number: AG019), caspase-1rabbit polyclonal antibody (catalog number: AF1681), Nrf2 rabbit polyclonal antibody (catalog number: AF7623), NF-κB p65 rabbit polyclonal antibody (catalog number: AF5875), HO-1rabbit polyclonal antibody (catalog number: AF1333), NLRP3 rabbit monoclonal antibody (catalog number: AF2155), HRP-labeled goat anti-rabbit IgG (H + L) (catalog number: A0208) and HRP-labeled goat anti-mouse IgG (H + L) (catalog number: A0216). Original Western Blot figures in Figure S2.

Mv.1.Lu cells were cultivated on cover glasses in 24-well plates, followed by infection with PS or CDV₃ at an MOI of 2 when the cells reached ~70% confluence. The mock-infected cells were treated with PBS as a negative control. Next, at 24 hpi, the cells were fixed with 4% paraformaldehyde and subsequently permeabilized with 0.1% Triton X-100. Further, the cells were incubated with an NF-κB P65 rabbit polyclonal antibody (Beyotime, China) and a mouse monoclonal antibody specific to CDV N protein and incubated with Cy3-labeled goat anti mouse IgG (Beyotime, China) and FITC-conjugated goat anti-rabbit IgG secondary antibody (ThermoFisher, U.S.A) prior to staining with DAPI. The fluorescent images were analyzed under confocal microscopy (Leica, Germany).