Ab95873

Cells were prepared at 4°C in RIPA buffer (P0013J, Beyotime, China) containing proteinase inhibitor cocktail (B14001, Bimake, Houston, TX, USA) and phosphatase inhibitor cocktail (B15001, Bimake). Proteins were separated on 4%–20% precast mini polyacrylamide gels (SurePAGE™, GenScript, Nanjing, China) and transferred to PVDF membranes. The membranes were incubated overnight at 4°C with anti-NEK7 (ab95873, Abcam), anti-GSDMD (ab219800, Abcam), anti-cleaved N-terminal GSDMD ab215203, Abcam), anti-NLRP3 (ab263899, Abcam), anti-cleavage caspase-1 (4199S, Cell Signaling Technology, Danvers, MA, USA), anti-ERK1/2 (9102S, Cell Signaling Technology), anti-p-ERK1/2 (9101S, Cell Signaling Technology), anti-α-SMA (A17910, ABclonal, Woburn, MA, USA), and anti-β-actin (66009-1-Ig, Proteintech, Rosemont, IL, USA). Membranes were then probed with appropriate secondary antibodies (Cell Signaling Technology). Immunoblot signals were detected by a Millipore chemical developer (Millipore Sigma, Burlington, MA, USA).

Cells were prepared at 4°C in RIPA buffer (P0013J, Beyotime, China) containing proteinase inhibitor cocktail(B14001, bimake, USA) and phosphatase inhibitor cocktail (B15001, bimake). Proteins were separated on 4–20% precast mini polyacrylamide gels (SurePAGE™, Genescript, China) and transferred to PVDF membrane. Membranes were incubated overnight at 4°C with anti-NEK7 (ab95873, Abcam), anti-β-actin (66009-1-Ig, Proteintech, USA), anti-β-Tubulin (A12289, Abclonal, China). Membranes were then probed with appropriate secondary antibodies (Cell Signaling Technology). Immunoblot signals were detected by a Millipore chemical developer (Millipore Sigma, Burlington, MA, USA).