Goat anti rabbit or anti mouse horse radish peroxidase labeled antibodies

Cells were washed with cold PBS and treated with RIPA buffer (89900; Thermo, Rockford, IL, USA) containing protease inhibitor cocktail (4693132001; Roche) and 1% phenylmethylsulfonyl fluoride PMSF (ST505; Beyotime, Shanghai, China). Proteins were quantified by Bradford assay and separated using a 12% SDS-PAGE gel prior to transferring onto a polyvinylidene difluoride (PVDF) membrane. Protein bands were visualized by enhanced chemiluminescence technique using SuperSignal West Pico chemiluminescent substrate (34577; Thermo, CA, USA). Densitometric quantification was performed using ImageJ v1.48 and the relative band intensity for each protein of interest was normalized against GAPDH. The primary antibodies used in this study included antibodies against GAPDH (60004-1-Ig; Proteintech, Wuhan, China), ACTIN (20536-1-AP; Proteintech), caspase-1 (#3866; Cell Signaling), α-catenin (#2131; Cell Signaling), β-catenin (#8480; Cell Signaling), α-SMA (#19245, Cell Signaling), vimentin (#5741; Cell Signaling). The goat anti-rabbit or anti-mouse horse radish peroxidase (HRP)-labeled antibodies were obtained from Proteintech.

Cells were washed with cold PBS and treated with RIPA buffer (Thermo, Rockford, IL) containing protease inhibitor cocktail (Roche) and 1% phenylmethylsulfonyl fluoride PMSF (Beyotime, Shanghai, China). Proteins were quantified by Bradford assay and separated in 12% SDS-PAGE prior to transferring onto polyvinylidene difluoride (PVDF) membrane. Protein bands were visualized by enhanced chemiluminescence technique using SuperSignal West Pico chemiluminescent substrate (Thermo, USA). Densitometric quantification was performed using ImageJ v1.48 and the relative band intensity for each protein of interest was normalized against β-actin. The primary antibodies used in this study included antibodies against β-actin (Proteintech, Wuhan, China), caspase-1 (Cell Signaling), EV71 VP1(Abnova), IL-1β (Proteintech), IL-18 (Proteintech), and NLRP3 (Biovector, Shanghai, China). The goat anti-rabbit or anti-mouse horse radish peroxidase (HRP)-labeled antibodies were obtained from Proteintech.